Nucleic Acid Amplification

Transcription

Nucleic Acid
Amplification
Product Guide
Quantify Gene Expression.
Save time and money with ready-to-order, predesigned
KiCqStart® Primers from Sigma® Life Science.
Bioconvenient.
Available through Sigma’s state-of-the-art gene search tool,
KiCqStart Primers are perfect for two-step and one-step
SYBR® Green I RT-qPCR. Together with our ReadyScript™ cDNA
Synthesis Mix and KiCqStart SYBR Green qPCR ReadyMix™ for
two-step reactions, Sigma ensures the success of every SYBR
qPCR assay — every time.
KiCqStart your assay now
sigma.com/ksprimers
©2012 Sigma-Aldrich Co. All rights reserved. SIGMA and SIGMA-ALDRICH are are trademarks of Sigma-Aldrich Co. LLC, registered in the US and other countries.
SYBR is a registered trademark of Molecular Probes, Inc. KiCqStart is a registered trademark of Sigma-Aldrich Co. LLC. OligoArchitect ReadyMix, ReadyScript, and
Where bio begins are trademarks of Sigma-Aldrich Co. LLC.
Table of Contents
Quick Reference Guides
2 Post-reaction Purification
Routine Amplification
4 Related PCR Reagents
Enzymes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Genomic DNA Amplification. . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
ReadyMixes and SuperPaks . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Long and Accurate PCR
7
Enzymes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Specialty Enzymes
8
Hot Start PCR
Enzymes. . .
ReadyMixes
CleanAmp™
Reagents . .
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Deoxynucleotides.
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RT-PCR
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Enzymes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Related Products for RT-PCR . . . . . . . . . . . . . . . . . . . . . . . . 16
Real-Time PCR
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Standard qPCR Reagents . . . . . . . . . . .
SYBR® Green . . . . . . . . . . . . . . . . . . . . . .
Probe-based . . . . . . . . . . . . . . . . . . . . . .
Fast qPCR Reagents . . . . . . . . . . . . . . .
SYBR® Green . . . . . . . . . . . . . . . . . . . . . .
Probe-based . . . . . . . . . . . . . . . . . . . . . .
MicroRNA RT-qPCR Assays. . . . . . . . . .
MystiCq® MicroRNA qPCR Primers . . . . .
Related Products for Real-time PCR. . .
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Whole Genome Amplification
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Whole Transcriptome Amplification
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Extract-N-Amp Kits
Blood Kits. . . . . . . . .
Plant. . . . . . . . . . . . .
Tissue . . . . . . . . . . . .
Seed. . . . . . . . . . . . .
SYBR® Green Tissue .
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Human Genomic DNA . . . . . . . . . . . . . . .
Deoxynucleotides . . . . . . . . . . . . . . . . . .
Deoxynucleotide Sets . . . . . . . . . . . . . . . . .
Individual Deoxynucleotides . . . . . . . . . . . .
Deoxynucleotide Mixes . . . . . . . . . . . . . . . .
PCR Buffers . . . . . . . . . . . . . . . . . . . . . . .
Extract-N-Amp™ ReadyMixes . . . . . . . . . .
SYBR® Green Extract-N-Amp™ ReadyMix .
PCR Optimizing Reagents . . . . . . . . . . . .
Post-reaction Analysis Reagents . . . . . . .
DNA Ladder Markers . . . . . . . . . . . . . . . . . .
RNA Markers . . . . . . . . . . . . . . . . . . . . . . . .
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PCR Accessories
Thermal Cyclers . . . . . . . . . . . . . . . . . .
Plates . . . . . . . . . . . . . . . . . . . . . . . . . .
Plate Seals . . . . . . . . . . . . . . . . . . . . . .
ThermalSeal® Films for Real-time PCR .
PurePak PCR Microtubes . . . . . . . . . . .
PCR Microtubes . . . . . . . . . . . . . . . . . .
48
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Sigma® Custom Products
The Sigma Advantage . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Expertise of Our Scientists. . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Outstanding Customer Service & Technical Support . . . . . . . .
Our Commitment to Quality. . . . . . . . . . . . . . . . . . . . . . . . . . .
Global Manufacturing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
OEM and Contract Manufacturing . . . . . . . . . . . . . . . . . . . . . .
Custom Oligonucleotides. . . . . . . . . . . . . . . . . . . . . . . . . . .
DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Primers Made Easy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Oligos in Plates for High-throughput Applications . . . . . . . . .
iScale Oligos™ DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
qPCR Probes and Assay Design Services. . . . . . . . . . . . . . .
Quantitative Real-time PCR Probes . . . . . . . . . . . . . . . . . . . . . .
Assay Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Trademark Index
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sigma.com
PCR Quick Reference Guide
Product Description
Routine PCR Products
Taq DNA Polymerase
Taq DNA Polymerase without MgCl2
REDTaq® DNA Polymerase
REDTaq® DNA Polymerase SuperPak™
ReadyMix™ Taq PCR Reaction Mix
REDTaq® ReadyMix™ PCR Reaction Mix with MgCl2
Long and Accurate PCR Products
AccuTaq™ LA DNA Polymerase
REDAccuTaq® LA DNA Polymerase
Hot Start PCR Products
JumpStart™ Taq DNA Polymerase
JumpStart™ Taq DNA Polymerase without MgCl2
JumpStart™ REDTaq® DNA Polymerase
JumpStart™ Taq ReadyMix™
JumpStart™ REDTaq® ReadyMix PCR Reaction Mix
JumpStart™ REDTaq® ReadyMix For High Throughput
PCR
JumpStart™ AccuTaq™ LA DNA Polymerase
JumpStart™ REDAccuTaq™ LA DNA Polymerase
JumpStart™ Taq Antibody
CleanAmp™ dNTPs
Easy MgCl2
Optimization
Direct
LoadA
Assembled
Master MixB
Proofreading
Enzyme with 3′→5′
Exonuclease Activity
Fidelity
Compared to
Standard TaqC
Amplification
Length (kb)D
1×
0.1 to >3 (5)
1×
0.1 to >3 (5)
Page
Cat. No.
4
D1806
4
D4545
4
D4309
✓
1×
0.1 to >3 (5)
6
D6063
✓
1×
0.1 to >3 (5)
5
P4600
✓
1×
0.1 to >3 (5)
6
R2523
✓
1×
0.1 to >3 (5)
7
D8045
✓
up to 6.5×
0.1 to >20 (40)
7
D4812
✓
up to 6.5×
0.1 to >20 (40)
9
D9307
1×
0.1 to >3 (10)
9
D4184
1×
0.1 to >3 (10)
10
D8187
1×
0.1 to >3 (10)
12
P2893
✓
1×
0.1 to >3 (10)
12
P0982
✓
✓
1×
0.1 to >3 (10)
12
P1107
✓
✓
1×
0.1 to >3 (10)
11
D5809
up to 6.5×
0.1 to >20 (40)
11
D1313
up to 6.5×
0.1 to >20 (40)
13
A7721
13
DNTPCA1
DNTPCA2
DNTPCA10
✓
✓
✓
✓
✓
✓
Includes
Separate
dNTP MixE
✓
A. REDTaq® and REDAccuTaq® products contain an inert red dye. The dye provides visual confirmation that the Polymerase has been added to the reaction and mixing is complete. Aliquots from the PCR can be
directly loaded onto the gel without adding loading buffers or tracking dyes. The dye has no effect on automated DNA sequencing, ligation, transformation or other downstream applications.
B. Each ReadyMix™ is conveniently supplied at 2× concentration and prepared using the indicated thermostable DNA Polymerase, ultrapure 99%+ dNTPs and high quality molecular biology reagents. To prepare a 50
μL PCR reaction, add 25 μL of the appropriate ReadyMix™ to 25 μL of water containing primers and template.
C. Fidelity compared to Taq DNA Polymerase.
D. Two values are provided for the indicated amplification length. The range provided refers to the average length achieved from complex genomic targets, while the number in parentheses refers to lengths routinely
achieved with less complicated targets such as plasmid or lambda phage DNA.
E. SuperPak™ convenient packages containing our high quality REDTaq® DNA Polymerase, 10 mM ultrapure dNTP mix, and 10× reaction buffer with or without MgCl2.
Quick Reference Guides
Sigma's Latest Mixes for Quantitative PCR
Sigma's Legacy Mixes for Quantitative PCR
These Sigma mixes offer the convenience of being pre-formulated with the
proper amount of passive reference dye, should your real-time PCR instrument
require it. Please select the proper formulation based on your application (fast
qPCR or microRNA RT-qPCR) and your instrument. If you have any questions,
contact Technical Service (sigma.com/techservice).
These Sigma mixes offer the flexibility of being used on a variety of real-time
PCR instruments. A vial of ROX passive reference dye (100×, Cat. No. R4526) is
provided with each of these reagents so that it can be mixed in at the proper
concentration based on your platform. Select the proper final ROX
concentration based on your instrument. If you have any questions, contact
Technical Service (sigma.com/techservice).
Real-time PCR
Instrument Name
Applied Biosystems 5700
Applied Biosystems 7500 Fast
Applied Biosystems 7900HT Fast
Applied Biosystems 7900HT
Applied Biosystems StepOnePlus™
Applied Biosystems StepOne™
Applied Biosystems ViiA™ 7
Bio-Rad CFX384™
Bio-Rad CFX96™
Bio-Rad iCycler iQ®
Bio-Rad iQ™5
Bio-Rad MiniOpticon™
Bio-Rad MyiQ™
Bio-Rad/MJ Chromo4™
Bio-Rad/MJ Opticon 2
Bio-Rad/MJ Opticon™
Cepheid SmartCycler®
Eppendorf Mastercycler® ep realplex
Eppendorf Mastercycler® ep realplex2 S
Illumina Eco qPCR
Qiagen/Corbett Rotor-Gene® 3000
Qiagen/Corbett Rotor-Gene® Q
Roche LightCycler™ 480
Stratagene Mx3000P®
Stratagene Mx4000™
Choose the proper formula of KiCqStartA (KCQSXX) or
MystiCqB (MIRRMXX) for your instrument
KCQS00/
KCQS01/
KCQS02/
KCQS03/
MIRRM00C
MIRRM01D
MIRRM02E
MIRRM03F
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
A. KiCqStart SYBR Green qPCR ReadyMixes are designed for fast qPCR, but can be run in standard mode.
B. MystiCq microRNA SYBR Green ReadyMix is for use with MystiCq microRNA RT-qPCR Products.
C. These formulas do not contain a passive reference dye.
D. These formulas contain ROX at a concentration that is suitable for instruments that utilize paired
excitation/emission filters.
E. These formulas contain ROX at a concentration that is suitable for instruments that utilize a single
excitation filter with multiple emission filters.
F. These formulas contain fluorescein at the proper concentration for certain Bio-Rad instruments.
S4438
SYBR® Green JumpStart™ Taq ReadyMix™
S5193
SYBR® Green JumpStart™ Taq ReadyMix™, without MgCl2
QR0100
SYBR® Green Quantitative RT-PCR Kit
D7440
JumpStart™ Taq ReadyMix™ for Quantitative PCR
QR0200
Quantitative RT-PCR ReadyMix™
L6544
LuminoCt® SYBR® Green qPCR ReadyMix™
L6669
LuminoCt® qPCR ReadyMix™
Real-time PCR
Instrument Name
Applied Biosystems 7500 Fast
Applied Biosystems 7900HT Fast
Applied Biosystems 7900HT
Applied Biosystems StepOnePlus™
Applied Biosystems StepOne™
Applied Biosystems ViiA™ 7
Bio-Rad CFX384™
Bio-Rad CFX96™
Bio-Rad iCycler iQ®
Bio-Rad iQ™5
Bio-Rad MiniOpticon™
Bio-Rad MyiQ™
Bio-Rad/MJ Chromo4™
Bio-Rad/MJ Opticon 2
Bio-Rad/MJ Opticon™
Cepheid SmartCycler®
Eppendorf Mastercycler® ep realplex
Eppendorf Mastercycler® ep realplex2 S
Illumina Eco qPCR
Qiagen/Corbett Rotor-Gene® Q
Roche LightCycler™ 480
Stratagene Mx4000™
For Cat. Nos. S4438, S5193, QR0100, L6669, D7440,
QR0200, L6544, dilute ROX (R4526) to final concentration
based on your instrument below
Final ROX
Final ROX
No ROX
Concentration 1× Concentration 0.1×
Needed
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
See Note A
See Note A
✓
See Note A
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
✓
See Note B
✓
See Note B
✓
See Note B
Notes
A. For Bio-Rad iCycler iQ®, iQ™ 5 and MyiQ™ systems, use fluorescein dye as instead of ROX
recommended by the manufacturer.
B. Stratagene platforms have the option of not utilizing a normalization dye, but it is recommended.
3
4
sigma.com
Enzymes
Taq DNA Polymerase from Thermus aquaticus
Taq DNA Polymerase is a specialized thermostable enzyme isolated from the
thermophilic bacterium Thermus aquaticus. The recombinant form of this
enzyme is expressed in E. coli. This 94 kDa protein shows no detectable levels
of contaminating endonucleases or exonucleases by SDS-PAGE. It has both
5′→3′ polymerase and exonuclease activity.
REDTaq allows quick recognition and confirms appropriate mixing.
Vial 1 contains no REDTaq, vial 2 has 2.5 units of REDTaq added to a 50 μL reaction volume, and vial 3
shows a REDTaq PCR reaction solution thoroughly mixed.
Taq DNA Polymerase from Thermus aquaticus is a thermostable DNA
polymerase that is used for the DNA polymerase chain reaction (PCR) in order
to amplify DNA sequences.
Taq DNA polymerase comes with the choice of an optimized 10× reaction
buffer including MgCl2 or a 10× reaction buffer without MgCl2 plus a separate
tube of MgCl2 for titration. The latter option may be necessary to determine
optimal conditions for amplification.
One unit will incorporate 10 nmol of total dNTPs into acid-precipitable DNA in
30 min at 74 °C.
concentration ....................................................................................................................................................... 5 units/μL
 with 10× PCR reaction buffer containing MgCl2
Taq DNA Polymerase with 10× reaction buffer containing MgCl2
D1806-250UN
D1806-20X250UN
D1806-1.5KU
D1806-10X1.5KU
D1806-5KU
No loading buffers or tracking dyes required with Sigma REDTaq DNA Polymerase.
Samples may be added directly to an agarose gel after PCR without the addition of a loading buffer
or tracking dye. The dye in REDTaq acts as a tracking dye migrating at approximately the same rate as a
125 bp fragment.
250 units
7 kb
20 × 250 units
1500 units
3 kb
10 × 1500 units
2 kb
5000 units
 with 10× PCR reaction buffer without MgCl2
1 kb
Taq DNA Polymerase with 10× reaction buffer without MgCl2. Includes a
separate tube of 25 mM MgCl2
D4545-50UN
50 units
D4545-250UN
250 units
D4545-20X250UN
20 × 250 units
Same great performance as standard Taq.
Comparison of yield for 1, 2, 3, and 7 kb DNA fragments using REDTaq (R) and standard Taq (T) DNA
polymerases under identical PCR cycling conditions.
D4545-1.5KU
1500 units
 Taq for routine PCR with inert dye, 10X buffer included
D4545-5KU-TAQ
5000 units
Provided with 10X reaction buffer containing MgCl2
D4545-5KU
5000 units
D4309-50UN
50 units
D4309-250UN
250 units
REDTaq® DNA Polymerase
Features and Benefits
• Same great performance as Taq DNA Polymerase in a more convenient
format for high throughput applications.
• Visual confirmation that not only has the enzyme been added, but that
proper mixing has occurred.
• No additional loading dyes are necessary. An aliquot can be taken directly
from the reaction and loaded onto an agarose gel for electrophoresis.
One unit incorporates 10 nmol of total dNTPs into acid-precipitable DNA
in 30 min at 74 °C.
concentration ......................................................................................................................................................... 1 unit/μL
D4309-1KU
1000 units
D4309-2.5KU
2500 units
10× REDTaq® PCR Reaction Buffer
 To be used with REDTaq® DNA Polymerase
10x REDTaq® PCR Reaction Buffer is a polymerase chain reaction buffer for use
with REDTaq® DNA Polymerase.
B5926-1.5ML
B5926-5ML
1.5 mL
5 mL
Enzymes: Genomic DNA Amplification
ReadyMixes and SuperPaks
Genomic DNA Amplification
ReadyMix™ Taq PCR Reaction Mix
REDTaq® Genomic DNA Polymerase
• Enhanced amplification on genomic and difficult DNA templates
• Same great performance as Taq DNA Polymerase in a more convenient
format for high throughput applications
• Quick recognition and confirmation of appropriate mixing
• No loading buffers or tracking dyes necessary. Sample can be taken directly
from reaction and loaded onto an agarose gel
ReadyMix™ Taq PCR Reaction Mix is a prepared solution containing everything
needed for a PCR reaction except the specific primers and template. The mix
includes Sigma′s high quality Taq DNA Polymerase, 99% pure deoxynucleotides and buffer in a 2× optimized reaction concentrate. For reaction set-up,
add the ReadyMix (25 μL) to the primers, template and water (total volume
50 μL). Using ReadyMix Taq PCR Reaction Mix reduces pipetting steps and risk
of contamination. This saves time and reduces errors while still providing the
great performance of Sigma′s Taq DNA Polymerase.
One unit incorporates 10 nmol of total dNTPs into acid precipitable DNA
in 30 min. at 74°C.
• Amplifies targets up to 7 kb in length
• ReadyMix Taq PCR Reaction Mix is provided either with MagCI2
• ReadyMix Taq PCR Reaction Mix offers the same great performance as Taq
DNA Polymerase in a more convenient formulation
• REDTaq® ReadyMix PCR Reaction Mix combines all the advantages of the
ReadyMix with the added convenience of a direct load system. After PCR, an
aliquot can be removed from the reaction and loaded directly onto an
agarose gel with no loading dye additions, making it ideal for high
throughput applications
M
1
2
3
4
M
Supplied with a vial of PCR grade water for dilution.
in 30 min. at 74 °C.
concentration .......................................................................... 1.5 units/reaction (50 μL reaction volume)
Higher yields from genomic templates with REDTaq Genomic DNA Polymerase.
PCR reactions were set up using 1 μL of mouse genomic DNA and 1 unit of polymerase. The resulting
amplicon is a specific 1181 bp fragment. Each sample was prepared in duplicate, and conditions for both
sets were identical with the exception of the enzyme used.
Lanes
M. 1 kb DNA marker
1 and 2. REDTaq DNA Polymerase
3 and 4. REDTaq Genomic DNA Polymerase
7 kb
4.5 kb
3 kb
2 kb
1 kb
 with MgCl2
Includes 10× PCR Reaction Buffer
D8312-250UN
M
250 units
D8312-1KU
1000 units
D8312-2.5KU
2500 units
2
3
4
5
M
Excellent amplification with ReadyMix Taq PCR Reaction Mix.
Amplification of 1, 2, 3, and 7 kb fragments and a 4.5 kb human genomic DNA using ReadyMix Taq PCR
Reaction Mix.
 with MgCl2
 without MgCl2
Includes 10× PCR reaction buffer without MgCl2 and a separate tube of 25 mM
MgCl2
D2812-250UN
1
250 units
D2812-1KU
1000 units
D2812-2.5KU
2500 units
Default reaction volume is 50 μL
100RXN is packaged as 1 X 2.5 mL
P4600-100RXN
100 reactions
5
6
sigma.com
REDTaq® ReadyMix™ PCR Reaction Mix
REDTaq® SuperPak™ DNA Polymerase
 Complete PCR reagent with standard Taq DNA Polymerase and inert dye
 Taq for routine PCR with inert dye; with 10X buffer & dNTP mix
REDTaq® SuperPak™ DNA Polymerase is used for PCR DNA amplification
methods. It contains the high visibility REDTaq® DNA polymerase and other
reagents needed for PCR reactions.
20RXN is packaged as 1 X 500 μL
R2523-20RXN
20 reactions
R2523-100RXN
100 reactions
Components
REDTaq Genomic DNA polymerase (1 unit/μL)
10 mM Deoxynucleotide mix
10× PCR buffer
D6063-50UN
50 units
D6063-250UN
250 units
Oligos For Your Application
PCR, cloning, sequencing and more.
Bioflexible.
Choose Sigma custom DNA and RNA oligos, and get the
performance you need from the name you trust.
Oligos for yor application, sigma.com/oligos
Enzymes
Enzymes
AccuTaq™ LA DNA Polymerase
AccuTaq LA DNA polymerase is an optimized blend of Sigma′s high quality
Taq DNA polymerase and a small amount of an additional polymerase that
exhibits 3′→5′ exonuclease or proofreading activity. By blending the Taq with
the right amount of this proofreading enzyme, misincorporation errors are
corrected, producing PCR amplicons that are longer and more accurate.
• Increased fidelity, up to 6.5× that of Taq DNA polymerase
• Efficiently and accurately produce amplicons up to 22 kb on genomic
templates and up to 40 kb on less complex templates such as lambda or
bacterial DNA.
The enzyme is provided with an optimized 10× reaction buffer for enhanced
amplification of complex templates. A separate vial of DMSO is included.
Addition of DMSO in the reaction at a final concentration of 1-4% may increase
yield and improve reliability of the system with some complex PCR targets.
in 30 min. at 74°C.
 High fidelity Taq enzyme, with 10X buffer & DMSO
AccuTaq™ LA DNA Polymerase is utilized to amplify DNA sequences including
genomic targets larger than 20 kb, as a result of a mixture of high quality Taq
polymerase with a proofreading polymerase.
concentration ....................................................................................................................................................... 5 units/μL
1
2
3
4
5
6
7
8
9
10 11 12
13 14 15 16
5
6
7
8
and
and
and
and
13.
14.
15.
16.
9 kb
10 kb
12 kb
15 kb
D8045-125UN
125 units
D8045-500UN
500 units
D8045-1.5KU
1500 units
REDAccuTaq® LA DNA Polymerase
REDAccuTaq® LA DNA polymerase is a blend of Sigma′s high quality Taq DNA
polymerase, a small amount of an additional polymerase that exhibits 3′→5′
exonuclease or proofreading activity, and our inert red dye. By blending the
Taq with the right amount of this proofreading enzyme, misincorporation
errors are corrected, producing PCR amplicons that are longer and more
accurate.
REDAccuTaq LA DNA polymerase allows for quick recognition in high
throughput applications as well as direct loading of amplification products
onto agarose gels for electrophoresis. The inert red dye has no effect on
automated sequencing, restriction enzyme digestion, ligation, or other
downstream manipulations. However, the PCR product is easily separated from
the dye by standard purification methods.
• Increased fidelity, up to 6.5× that of Taq DNA polymerase
• Efficiently and accurately produce amplicons up to 22 kb on genomic
templates and up to 40 kb on less complex templates such as lambda or
bacterial DNA
• Visual confirmation that enzyme has been added, and proper mixing has
occurred
• No additional loading dyes needed. A post-reaction aliquot can be directly
used on an agarose gel for electrophoresis
The enzyme is provided with an optimized 10× reaction buffer for enhanced
amplification of complex templates.
 High fidelity Taq with inert dye, 10X buffer included
D4812-250UN
250 units
AccuTaq™ LA 10× Buffer
 10X Buffer for long and accurate PCR
Longer products with improved fidelity with AccuTaq LA DNA polymerase.
Amplification of 2.5, 7, 9, 10, 12 and 15 kb fragments of lambda DNA. 2.5 μg of DNA was amplified using
2.5 units of enzyme. The resulting products (6% of total reaction) were electrophoresed on a 0.8%
agarose gel. Lanes 3-8 contained Taq DNA polymerase and lanes 11-16 contained AccuTaq LA DNA
Polymerase.
Lanes
1 and 9. 1 kb DNA Marker
2 and 10. Lambda Hind III DNA Marker
3 and 11. 2.5 kb
4 and 12. 7 kb
10× Buffer for AccuTaq LA DNA Polymerase, product code D8045 and D4812
vial = 0.5 mL
B0174-1VL
1 vial
B0174-.5ML
0.5 mL
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sigma.com
Specialty Enzymes
MTP™ Taq DNA Polymerase
Restorase® DNA Polymerase with 10× Reaction Buffer
MTP Taq DNA Polymerase is a recombinant thermostable enzyme from
Thermus aquaticus expressed in E. coli and purified using a proprietary process
that minimizes levels of contaminating DNA. The enzyme has 5′-3′ DNA
polymerase and exonuclease activities, is approximately 95 kD by SDS-PAGE,
and has no detectable endonuclease or 3′-5′ exonuclease activities.
Contaminating DNA present in most other polymerase preparations often
precludes or obscures the accurate interpretation of results, especially when
targeting conserved sequences (e.g. bacterial 16S rRNA region).
While MTP Taq is a high-quality, low-contaminant DNA polymerase for reliable
PCR amplification, DNA contaminants can be introduced into PCR through a
number of other reagents. To further minimize the risk of contaminant DNA
during PCR, we include 10× MTP Taq buffer (Sigma product M 9943) with each
tube of MTP Taq DNA Polymerase. Each lot of MTP Taq and 10× MTP Taq
buffer undergoes the same strict quality control testing to ensure the absence
of contaminating DNA. To prevent false positive PCR results, only DNA-free
reagents should be used in PCR reactions with MTP Taq DNA Polymerase.
 Taq DNA Polymerase, free of DNA contaminants
D7442-250UN
250 units
D7442-1500UN
1500 units
10X MTP™ Taq Buffer
DNA contaminants can be introduced into PCR through a number of reagents.
To minimize the risk of contaminant DNA during PCR, we offer 10x MTP Taq
buffer to be used with MTP Taq DNA Polymerase (Sigma product D7442). Each
lot of MTP Taq buffer undergoes strict quality control testing to ensure the
absence of contaminating DNA. To prevent false positive PCR results, only
DNA-free reagents should be used in PCR reactions with MTP Taq DNA
polymerase.
 10X buffer, free of DNA contaminants; use with MTP Taq DNA Polymerase
M9943-1.5ML
1.5 mL
Restorase DNA Polymerase with 10× Reaction Buffer combines Sigma′s long
and accurate enzyme technology with a small amount of DNA repair enzyme.
The optimized blend will initiate the repair and further amplification of
damaged DNA templates greater than 800 bp. Restorase has also been shown
to increase yield on undamaged DNA templates.
• Reliable amplification of damaged DNA
• Increase Yield
• Can repair 3′ bungs, nicks, and abasic sites
The enzyme is provided with an optimized 10× reaction buffer provided
as 1 vial/250 units.
 Enzyme blend for PCR amplification of damaged DNA
concentration ................................................................................................................................................... 2.5 units/μL
R1028-20RXN
20 reactions
R1028-50RXN
50 reactions
R1028-200RXN
200 reactions
Hot Start PCR
Enzymes
Hot Start PCR
Enzymes
JumpStart™ Taq DNA Polymerase
Sigma′s JumpStart Taq DNA Polymerase is an antibody-inactivated hot-start
enzyme designed to minimize non-specific amplification while increasing
target yield. Once the reaction temperature reaches 70°C, Taq DNA polymerase
activity is restored and the resulting PCR exhibits a higher specificity and yield.
This antibody-enzyme complex allows for easy and convenient set-up with less
contamination risk than manual hot-start techniques. The enzyme may also be
included in the master mix preparation resulting in more consistency from one
reaction to the next.
• Greater Specificity & Increased Target Yield. JumpStart Taq Polymerase, an
antibody inactivated hot start enzyme, is designed to minimize non-specific
amplification while increasing target yield & specificity
• Enhanced Sensitivity. JumpStart Taq DNA Polymerase provides superior
amplification regardless of template concentration
• Ultimate Convenience. Reduce set-up time and eliminate concerns
associated with manual or wax Hot Start methods
JumpStart Taq DNA Polymerase is provided with a 10× reaction buffer
available with and without MgCl2. The magnesium free 10× buffer also
includes a separate tube of 25 mM MgCl2 for optimization.
concentration ................................................................................................................................................... 2.5 units/μL
JumpStart Taq delivers better specificity
1.5 ng of total human genomic DNA was amplified with primers targeted to a 5 kb section of β-globin.
Reactions were formulated according to the supplier′s recommendations. Taq was activated per the
supplier′s recommendations.
 with MgCl2
Supplied with 10× reaction buffer containing 15 mM MgCl2
D9307-50UN
50 units
D9307-250UN
250 units
D9307-1.5KU
1500 units
 without MgCl2
Supplied with 10× reaction buffer without MgCl2. Includes a separate tube of
25 mM MgCl2
JumpStart Taq amplifies up to 9 kb
200 ng Lambda DNA was amplified with JumpStart Taq DNA Polymerase using recommended cycling
parameters.
D4184-50UN
50 units
D4184-250UN
250 units
D4184-1.5KU
1500 units
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sigma.com
JumpStart™ REDTaq® DNA Polymerase
JumpStart™ REDAccuTaq® LA DNA Polymerase
JumpStart REDTaq DNA Polymerase is Sigma′s high performance Taq DNA
Polymerase blended with JumpStart Taq antibody and an inert red dye tracer.
Extensive testing with a variety of primers and templates indicates that the
performance of JumpStart REDTaq DNA Polymerase is equivalent to, or better
than, that of standard Taq polymerase.
The inert red dye does not affect automated sequencing, restriction enzyme
digestion, ligation or other downstream applications. The PCR product can be
easily separated from the dye by standard purification methods.
• Prevents amplification of nonspecific products, resulting in increased
efficiency and higher yields of the desired sequence
• No lengthy activation step is required for enzyme activity to be restored
• Visual confirmation that the enzyme has been added and that proper
component mixing of the reaction has occurred
• Samples can be loaded directly onto an agarose gel for electrophoresis with
no loading dye additions
The enzyme is provided with an optimized 10× reaction buffer.
 Hot-start high fidelity Taq enzyme with inert dye, 10X buffer included
–Ab
M
+Ab
1
2
3
4
5
6 7
8
4 ng: JumpStart REDTaq DNA Polymerase improves yield and specificity across a broad range of
template
Four nanograms of template DNA was amplified under standard PCR conditions (lanes 1−4) and Hot
Start conditions using JumpStart REDTaq (lanes 5−8). Higher yields using JumpStart REDTaq are evident
in lanes 5−8, as well as reduced non-specific products.
–Ab
JumpStart REDAccuTaq LA DNA Polymerase is a unique enzyme blend that is
capable of generating long PCR fragments, from 0.25 kb to 40 kb, with high
fidelity, increased specificity and yield. JumpStart REDAccuTaq DNA polymerase combines Sigma′s AccuTaq LA DNA polymerase and JumpStart Taq
antibody with an inert red dye. This specially formulated hot start enzyme mix
achieves greater yields, enhances sensitivity and results in higher fidelity (6.5×)
in comparison to standard Taq or other Long and Accurate enzyme blends. Its
high fidelity makes it the enzyme of choice when performing amplifications
where a low error frequency is critical, such as in RT-PCR and cloning.
JumpStart REDAccuTaq LA DNA Polymerase also contains the hot start
mechanism of the JumpStart Taq antibody. JumpStart Taq antibody is
designed to minimize non-specific amplification while increasing target yield.
Unlike other hot-start methods (i.e. chemical inactivation), JumpStart Taq
antibody does not require a pre-incubation step prior to cycling because
polymerase activity is fully restored during the first denaturation cycle of the
PCR reaction.
The inert red dye provides quick recognition and confirmation of appropriate
mixing. An aliquot of the samples (5-10 μL) may be loaded directly onto an
agarose gel following PCR. The red dye migrates slightly faster than
bromophenol blue at the same rate as a 125 base pair fragment.
The PCR product can be easily separated from the dye by standard purification
methods. The inert red dye has does not effect automated sequencing,
restriction enzyme digestion, ligation or other downstream applications.
• JumpStart REDAccuTaq LA DNA polymerase, an antibody inactivated hot
start enzyme, is designed to minimize non-specific amplification while
increasing target yield & specificity
• Up to 6.5X greater fidelity in comparison to Taq DNA polymerase making it
the ideal enzyme for multiplex PCR
• Produce amplicons up to 22 kb with genomic templates and up to 40 kb
with less complex templates such as lambda or bacterial genomic DNA
• Reduce set-up time and eliminate concerns associated with manual or wax
Hot Start methods
• Dye allows for quick visual confirmation that reagent has been added and
mixed properly
• Direct loading onto an agarose gel without additional dyes
Supplied with optimized 10× reaction buffer
+Ab
M 1
2
3
4
5
6
7
8
0.4 ng: JumpStart REDTaq DNA Polymerase improves yield and specificity across a broad range of
template
Standard PCR conditions (lanes 1−4) and Hot Start conditions using JumpStart REDTaq (lanes 5−8) were
used to amplify 0.4 ng template DNA. Using hot-start PCR, there is a marked decrease in non-specific
product formation as well as an overall increase in yield of the desired PCR product.
D8187-50UN
50 units
D8187-250UN
250 units
D8187-2.5KU
2500 units
Hot Start PCR
Enzymes
 Long and accurate hot-start Taq with inert dye, 10X buffer included
 Hot-start high fidelity Taq enzyme, 10X buffer included
concentration ...................................................................................................................................................... 2.5 unit/μL
1
2
3
4
5
6
7
8
9
10 11
12 13 14
15 16
Improved target yield with JumpStart™ REDAccuTaq® LA DNA Polymerase.
Amplification of 7, 9, 10, 12, 15 and 17.5 kb fragments of DNA. 2.5 μg of DNA was amplified using
2.5 units of enzyme. The resulting products (6% of total reaction) were electrophoresed on a 0.8%
agarose gel. Lanes 3−8 contained AccuTaq LA DNA polymerase and lanes 11−16 contained JumpStart
REDAccuTaq LA DNA Polymerase.
Lanes
1 and 9. 1 kb DNA Marker
2 and 10. Lambda Hind III DNA Marker
3 and 11. 7 kb
4 and 12. 9 kb
5 and 13. 10 kb
6 and 14. 12 kb
7 and 15. 15 kb
8 and 16. 17.5 kb
D1313-50UN
50 units
D1313-250UN
250 units
M
1
2
3
4
5
JumpStart AccuTaq LA and REDAccuTaq LA DNA both outfperform the competition.
Long and accurate hot start enzymes were used to amplify a 5 kb fragment starting with 25 ng of total
human genomic DNA. All reactions were performed according to the manufacturer′s specifications.
Lanes
M. Wide Range DNA marker
1. JumpStart AccuTaq LA
2. JumpStart REDAccuTaq LA
3. Supplier S
4. Supplier I, enzyme P
5. Supplier I, enzyme HF
D5809-125UN
125 units
D5809-500UN
500 units
JumpStart™ AccuTaq™ LA DNA Polymerase
JumpStart AccuTaq LA DNA Polymerase is a combination of AccuTaq LA DNA
Polymerase and a Taq-directed antibody. JumpStart Taq antibody reversibly
binds to the AccuTaq LA DNA Polymerase, inactivating it at room temperature.
The increased temperature of the first denaturation cycle causes the complex
to dissociate, restoring the enzyme activity. This hot start mechanism provides
increased specificity and higher target yield in comparison to standard
amplification. JumpStart AccuTaq LA DNA Polymerase can generate long
products with higher fidelity (up to 6.5× of Taq DNA polymerase).
• JumpStart AccuTaq LA DNA polymerase, an antibody inactivated hot start
enzyme, is designed to minimize non-specific amplification while increasing
target yield & specificity
• Up to 6.5× greater fidelity in comparison to Taq DNA polymerase making it
the enzyme of choice for multiplex PCR
• Produce amplicons up to 22 kb with genomic templates and up to 40 kb
with less complex templates such as lambda or bacterial genomic DNA
hot-start methods
Supplied with 10× reaction buffer.
One unit incorporates 10 nmol of total dNTPs into acid-precipitable DNA in
30 min. at 74 °C.
ReadyMixes
JumpStart™ Taq ReadyMix™
JumpStart Taq ReadyMix is a prepared solution combining the performance
benefits of hot start PCR with the convenience of a ReadyMix. The mix includes
JumpStart Taq DNA polymerase, 99% pure deoxynucleotides and buffer in a
2× optimized reaction concentrate. JumpStart Taq Polymerase is an antibodyinactivated hot-start enzyme designed to minimize non-specific amplification
while increasing target yield. Unlike other hot-start methods (i.e. chemical
inactivation), JumpStart Taq polymerase does not require a pre-incubation
step prior to cycling because polymerase activity is fully restored during the
first denaturation cycle of the PCR reaction. The hot start mechanism allows for
room temperature set up, making it ideal for high throughput applications. To
prepare a 50 μL reaction, simply add 25 μL of ReadyMix to template, primers
and water for a final reaction volume of 50 μL.
• Reduce set-up time, eliminate contamination concerns, and avoid long
reactivation steps with JumpStart Readymixes
• JumpStart Taq antibody allows for room temperature set up of reactions
making it ideal for high throughput applications
• Prevents amplification of non-specific products and primer dimers,
increasing target yields
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sigma.com
M 1 2 3 4 5 6 M 7 8 9 10 11 12
400RXN is packaged as 1 X 10 mL
 Complete optimized reagent for hot-start PCR at 2X concentration
P2893-100RXN
100 reactions
P2893-400RXN
400 reactions
JumpStart™ REDTaq® ReadyMix™ Reaction Mix
JumpStart REDTaq ReadyMix PCR Reaction Mix is a prepared solution
combining the performance benefits of hot start PCR with the dual
convenience of Sigma′s ReadyMix and REDTaq. The mix includes Sigma′s
JumpStart Taq DNA Polymerase, 99% pure deoxynucleotides, buffer, and an
inert red dye in a 2× optimized reaction concentrate. Add 25 μL of the
ReadyMix to primers, template, and water to a final reaction volume of 50 μL.
JumpStart Taq antibody in the reaction mix inactivates the Taq DNA
polymerase at room temperature. During the first denaturation step of PCR the
complex dissociates and polymerase activity is fully restored.
The red dye allows quick visual confirmation the enzyme has been added and
properly mixed. After PCR, an aliquot may be loaded directly onto an agarose
gel without the addition of loading buffers. The inert red dye does not effect
automated sequencing, restriction enzyme digestion, ligation, or other
downstream manipulations. The PCR product is easily separated from the dye
by standard purification methods if desired.
Using JumpStart ReadyMix reduces pipetting steps and risk of contamination.
The hot start mechanism allows for room temperature set up, making it the
ideal for high throughput applications.
• JumpStart Taq DNA polymerase, an antibody inactivated hot start enzyme, is
designed to minimize non-specific amplification while increasing target yield
& specificity
• JumpStart Taq DNA polymerase provides superior amplification regardless of
template concentration
• REDTaq JumpStart ReadyMix reduces pipetting steps and risk of contamination. The hot start mechanism allows for room temperature set up,
making it the ideal for high throughput applications
• Inert red dye allows for easy verification that reagent has been mixed
properly
• Other loading dyes not necessary. Aliquots may be directly loaded onto an
agarose gel after the reaction
Exceptional performance with JumpStart REDTaq ReadyMix Hot Start PCR in the convenience of a
ReadyMix.
200 ng Lambda phage DNA was amplified with Sigma’s JumpStart REDTaq ReadyMix (odd numbered
lanes) and Competitor I’s Direct Load ReadyMix (even numbered lanes). Taq was activated per the
supplier’s recommendations.
P0982-20RXN
20 reactions
P0982-100RXN
100 reactions
P0982-800RXN
800 reactions
 for High-throughput PCR of complex templates
JumpStart REDTaq ReadyMix PCR Reaction Mix for High Throughput PCR is
formulated with REDTaq Genomic DNA polymerase for amplification of more
complex templates and genomic templates. This mix contains optimized
enzyme and dye concentrations to provide increased length and yield on the
more difficult templates.
concentration ....................................................................... 0.75 units/reaction (50 μL reaction volume)
P1107-100RXN
100 reactions
P1107-400RXN
400 reactions
CleanAmp™ Deoxynucleotides
CleanAmp™ dNTP
CleanAmp dNTPs, are a product of TriLink BioTechnologies, Inc. CleanAmp
dNTPs are modified nucleoside triphosphates that block DNA polymerase
nucleotide incorporation. CleanAmp dNTPs are activated by the initial heating
step and subsequent denaturing steps in typical hot start PCR cycling
conditions. This process limits the amount of activated dNTPs during each
cycle of PCR, allowing for more specific and efficient amplification of the
desired product and also reducing or completely avoiding mis-priming or
primer dimer formation. CleanAmp dNTPs provide a more affordable solution
than hot start enzymes for a variety of PCR-based applications.
Standard dNTPs
CleanAmp™ dNTPs
 for PCR
533 bp 715 bp 365 bp 653 bp
CleanAmp dNTPs demonstrate better yield and specificity than standard dNTPs for amplicons of varying
lengths.
Hot Start PCR
CleanAmp™ Deoxynucleotides
Standard dNTPs
CleanAmp™ dNTPs
 Modified dNTP set for hot-start PCR
650 bp
515 bp
388 bp
293 bp
214 bp
185 bp
114 bp
10 μmoles: 4 vials:
200 μL dATP at 50 mM in buffered glycine solution
200 μL dCTP at 50 mM in buffered glycine solution
200 μL dTTP at 50 mM in buffered glycine solution
200 μL dGTP at 50 mM in buffered glycine solution
DNTPCA10-1KT
1
7
1
1 kit
7
Number of Targets
CleanAmp dNTPs have been shown to effectively multiplex up to 7 different PCR targets without primer
dimer formation and also demonstrate improved yields for each assay when compared to standard
dNTPs.
Reagents
JumpStart™ Taq Antibody
The primary purpose of all hot start PCR methods is to prevent Taq DNA
polymerase activity prior to thermal cycling. Even if set-up is conducted on ice,
the Taq DNA polymerase remains active and may elongate unwanted
products such as misprimed or other non-specific events. Primer-dimer
interactions may also be amplified, which will reduce overall yield and
efficiency of desired products.
365 bp
Standard dNTPs
CleanAmp™ dATP, dTTP
with standard dCTP, dGTP
CleanAmp™ dNTP Mix
The CleanAmp dNTP Mix eliminates primer dimer formation, but similar improvements can be seen by
substituting standard dATP and dTTP with CleanAmp dATP and dTTP. The optimal base(s) to substitute
should be determined empirically for each assay.
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One way to prevent unwanted amplification products is to add JumpStart Taq
Antibody to the reaction. This efficient yet simple procedure takes only
10 minutes and effectively inactivates the Taq DNA polymerase until the first
denaturation cycle. Upon heating to 70 °C, the antibody dissociates and full
activity is restored to the Taq DNA polymerase for the remainder of the PCR.
Unlike chemically inactivated hot-start methods, no extended heating step is
required for reactivation with the use of JumpStart Taq Antibody.
JumpStart Taq antibody works effectively on a variety of commercially
available Taq DNA polymerases.
500 bp
• Minimize non-specific amplification while increasing target yield & specificity
hot-start methods.
JumpStart Taq Antibody, 1.1μg/1μL
Dilution Buffer provided in 1 mL vials
Standard dNTPs
Two units of Taq DNA polymerase are inactivated by 1 test of JumpStart Taq
Antibody.
CleanAmp™ dNTPs
CleanAmp dNTPs work well with a variety of DNA Polymerases to improve PCR performance.
 Hot-start dNTP mix for improved PCR
 Adds hot-start capabilities to any Taq DNA Polymerase
2 μmoles: 1 vial, 200 μL, each dNTP at 10 mM in 50 mM Glycine
10 μmoles: 1 vial, 1000 μL, each dNTP at 10 mM in 50 mM Glycine
DNTPCA1-2UMOL
2 μmol
DNTPCA1-10UMOL
10 μmol
 Modified dNTP set for hot-start PCR
2 μmoles: 4 vials:
40 μL dATP at 50 mM in buffered glycine solution
40 μL dCTP at 50 mM in buffered glycine solution
40 μL dTTP at 50 mM in buffered glycine solution
40 μL dGTP at 50 mM in buffered glycine solution
DNTPCA2-1KT
Supplied with dilution buffer.
1 kit
A7721-200TST
200 test
A7721-500TST
500 test
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sigma.com
RT-PCR
Enzymes
Enhanced Avian Reverse Transcriptase [eAMV™ RT]
M-MLV Reverse Transcriptase
Enhanced Avian Reverse Transcriptase (eAMV RT) is used to transcribe RNA into
DNA, and facilitates efficient mRNA template driven sysnthesis of cDNAs. This is
due to the abillity of this enhanced AMV-RT to transcribe large mRNA
templates, to transcribe through difficult secondary structures, and to detect
low abundance mRNAs by RT-PCR.
M-MLV (Moloney Murine Leukemia Virus) reverse transcriptase is a DNA
polymerase that uses single-stranded RNA, DNA, or an RNA-DNA hybrid (using
a primer) to synthesize a complementary DNA strand. M-MLV is used for the
preparation of cDNA libraries or for first strand cDNA synthesis for use in
RT-PCR reactions.
• Greater sensitivity for low abundance mRNA
• Unsurpassed transcription through difficult secondary structures at elevated
temperatures (up to 65°C)
• Efficient generation of full-length cDNA, up to 14.1 kb
The enzyme is purified from Escherichia coli expressing the pol gene of M-MLV
on a plasmid.
Provided with a vial of 10× reaction buffer.
 Moloney Murine Leukemia Virus enzyme & buffer for cDNA synthesis
One unit incorporates one nanomole of TMP into TCA precipitable material in
10 min using polyadenylic acid as template and oligo(dT)12-18 as a primer.
concentration .................................................................................................................................................. 200 units/μL
Supplied with 10× M-MLV reverse transcriptase buffer containing DTT.
One unit incorporates 1 nmol of TTP into acid precipitable material in 10 min.
at 37 °C using poly(A):oligo dT as a template:primer.
M1302-40KU
40000 units
 For reverse transcription at higher temperatures & rare mRNAs
concentration .................................................................................................................................................... 20 units/μL
Kits
ReadyScript® cDNA Synthesis Mix
ReadyScript cDNA Synthesis Mix is a sensitive and easy-to-use solution for twostep RT-PCR. This 5X concentrated mix provides all necessary components
(except RNA template) for first-strand synthesis including: buffer, dNTPs, MgCl2,
primers, RNase inhibitor protein, ReadyScript reverse transcriptase and
stabilizers. ReadyScript is a RNase H(+) derivative of MMLV reverse transcriptase, optimized for reliable cDNA synthesis over a wide dynamic range of
input RNA. The unique blend of oligo (dT) and random primers in the
ReadyScript cDNA Synthesis Mix works exceptionally well with a wide variety
of targets. It is optimized for the production of targets < 1kb in length.
ReadyScript cDNA Synthesis Mix produces excellent results in both real-time
and conventional RT-PCR.
eAMV™ RT efficiently reverse transcribes mRNA into cDNA up to 14.1 kb in size.
mRNA from HEK293 cells was reverse transcribed using eAMV Reverse Transcriptase to a known length
of 14.1 kb. The resulting cDNA was amplified using JumpStart REDTaq with PCR primer sets chosen at
different distances from the poly (A)+ tail resulting in the following PCR products. Successful
amplifications demonstrate the integrity of the cDNA up to 14 kb. Lanes 2-8 are primer sets for p619,
Lane 10 is actin and Lane 11 is GAPDH.
Lanes
1. Marker
2. 908 bp PCR product, 2,180 transcript size
9. Negative Control (No RT)
11. 452 bp PCR product, 880 transcript size
A4464-500UN
A4464-1KU
500 units
1000 units
• Convenient 5X master mix formula provides for quick and easy reaction set
up; just add RNA template
• Includes proprietary blend of randomers and oligodT primers and an RNAse
H+ derivative of Moloney murine leukemia virus (MMLV) reverse
transcriptase
• Allows for unbiased, accurate conversion of RNA to cDNA in just 40 minutes
• Can use up to 1 μg of RNA in a single 20 μL reaction; reactions can also be
scaled up
• Highly stable; can be stored at -20°C for up to 1 year; 4°C for up to one
month
 Complete reagent for first strand cDNA synthesis for RT-qPCR
RDRT-25RXN
25 reactions
RDRT-100RXN
100 reactions
RDRT-500RXN
500 reactions
RT-PCR
Kits
Enhanced Avian First Strand Synthesis Kit
eAMV™
Enhanced Avian First Strand Synthesis Kit utilizes a highly purified avian
myeloblastosis virus reverse transcriptase (eAMV-RT) that offers superior
performance in comparison to standard AMV-RT or standard Moloney murine
leukemia virus reverse transcriptase (MMLV-RT). This exceptionally robust
eAMV-RT has an enhanced ability to transcribe through difficult secondary
structure at elevated temperatures (up to 65 °C) making it the ideal enzyme for
producing high quality full-length cDNA from total RNA or poly(A)+ RNA.
One unit incorporates one nanomole of TMP into TCA-precipitable material in
10 minutes using polyadenylic acid as template and oligo(dT)12-18 as primer.
 Components for cDNA synthesis with enhanced AMV reverse transcriptase
1 kit sufficient for 50 reactions
Components
Enhanced Avian Reverse Transcriptase 1,000 units
10X Buffer for eAMVTM reverse transcriptase 1.5 mL
Deoxynucleotide mix 50 μL
Anchored oligo (dT)23 100 μL
Random nonamers 100 μL
Ribonuclease inhibitor 50 μL
PCR grade water 1.5 mL
STR1-1KT
1 2 3 4 5 6 7 8 9 10 11
Competitor M-MLV RT
RNase H-minus
1 2 3 4 5 6 7 8 9 10 11
Comparison of eAMV™ to M-MMLV RT RNase H- enzyme.
eAMV Reverse Transcriptase offers superior performance in length and yield over M-MLV RNase H-minus
enzymes when transcribing long cDNA. Different poly (A)+ RNA were used as a template in a two-step
RT-PCR.
Lanes
1. 1 kb Ladder
2 and 3. 2 kb Pol
4 and 5. 3.5 kb Pol
6 and 7. 5.3 kb TSC-2
8 and 9. 6.8 kb Pol
10 and 11. 8.9 kb APC
eAMV™
Standard AMV RT
1 kit
Enhanced Avian HS RT-PCR Kit
Reverse Transcriptase PCR (RT-PCR) is a powerful tool used to study gene
expression. The Enhanced Avian HS RT-PCR kit utilizes an enhanced avian
myeloblastosis virus reverse transcriptase (eAMV-RT) enzyme that offers
superior performance in comparison to standard AMV-RT and Moloney murine
leukemia virus reverse transcriptase (MMLV-RT). eAMV RT is an exceptionally
robust enzyme with an enhanced ability to transcribe through difficult
secondary structure at elevated temperatures (up to 65 °C) making it the ideal
enzyme for producing high quality full-length cDNA (up to 14.1 kb) from total
RNA or poly(A)+ RNA. JumpStart™ AccuTaq™ LA DNA polymerase mix is also
provided to eliminate non-specific amplification and increase specificity and
sensitivity. The combination of these two enzymes provides a quality system
that offers the versatility of a one-step or two-step RT-PCR protocol.
Procedures are provided for one-step and two-step RT-PCR reactions.
One-step: In a single tube, eAMV™ RT and AccuTaq LA act sequentially to first
produce cDNA and then immediately amplify by PCR. This provides quick,
sensitive analysis of RNA.
Two-step: Each reaction is individually optimized for greater yields with high
fidelity, when protocol requires multiple amplifications, or if maximum yield is
more important than maximum convenience.
eAMV™ RT demonstrates improved performance at higher temperatures.
RT-PCR was performed on 1.7 kb TMV transcript containing difficult secondary structure. The primer used
for the RT reaction is located in a region with extensive secondary structure, making reverse transcription
at an elevated temperature necessary.
15
sigma.com
7
8
9
10
11
12
Amplification Grade DNase I demonstrates lower RNase activity than that from
several leading molecular biology product suppliers.
13
706 bp
←
467 bp
Competitors
u
←
• Suitable for the elimination of DNA from RNA
• Minimal RNase activity available
• Optimized 10× reaction buffer and Stop Solution for complete inactivation
of DNase I
low abundance
M
6
W
5
Q
4
T
3
Same Order as Lanes 2–7
M-MLV RT
I
2
AMV-RT
E
1
eAMV™
C
i
Si
gm
a
R
M
C
16
medium abundance
eAMV™ RT outperforms competition when targeting low abundance genes.
RT-PCR was performed on human phopholipase A2, a low abundance RNA, (lanes 2-7) and human HPRT,
a medium abundance RNA (lanes 8-13), using eAMV RT, RNase H-reduced AMV, and RNase H-minus
M-MLV. Duplicate RT reactions were performed for each enzyme using 50 μg of HeLa poly (A)+ RNA and
reactions from each enzyme were pooled together before PCR. Two μL of cDNA was used for each PCR
reaction.
Lanes
1. DNA marker
2 and 3. RT-PCR using eAMV reverse transcriptase
4 and 5. RT-PCR using RNase H-reduced AMV-RT
6 and 7. RT-PCR using RNase H-minus M-MLV RT
8 thru 13. Same enzyme order as Lanes 2-7
 Flexible kit for one-step or two-step RT-PCR
 Amplification Grade
One unit completely digests 1 μg of plasmid DNA to oligonucleotides
Components
eAMV™ Reverse Transcriptase 2 x 1000U
JumpStart™ AccuTaq LA DNA Polymerase 2 x 125U
Random nonamers 100 μL
Anchored oligo(dT)23 primers 100 μL
10x reaction buffers 10 X AccuTaq Buffer
10 mM dNTP mix 10x PCR Buffer
RNase Inhibitor 100 μL
Nuclease-free water 4x1.5mL
HSRT100-1KT
Sigma Amplification Grade DNase I has the lowest RNase activity.
For Sigma DNase I, and for each competitor′s DNase I, the following assay was completed: 1 μg of a
1.9 kb in vitro transcription product was incubated with 1 unit of the respective DNase I at 37 °C for
1 hour and analyzed on a 1% agarose gel.
Cu = unincubated control (RNA in buffer without DNase, kept on ice).
Ci = incubated control (RNA in buffer without DNase, incubated at 37 °C for 1 hour).
Note: To determine the effectiveness of DNase I treatment, parallel PCR reactions should be run without
adding reverse transcriptase to check for amplification from contaminating DNA.
Components
DNase I 1,000 units
10X Reaction buffer 1 mL
Stop solution 1 mL
AMPD1-1KT
1 kit
Related Products for RT-PCR
DNase I
Deoxyribonuclease I (DNase I) is an endonuclease isolated from bovine
pancreas that digests double- and single-stranded DNA into oligo- and
mononucleotides. Using the Reaction Buffer provided, DNA is removed from
RNA preparations in a 15 minute digestion at room temperature. The DNase I
is then inactivated by heating with the Stop Solution. Heating also denatures
hairpins in the RNA, so the RNA can be used directly in reverse transcription.
Many commercial DNase I formulations are contaminated with residual
RNases. This RNase contamination can destroy or degrade valuable RNA
samples prior to reverse transcription. Laboratory comparisons have shown
that Sigma′s Amplification Grade DNase I demonstrates lower RNase activity
than that from several leading molecular biology product suppliers.
No current RNA isolation procedure removes 100% of the DNA. Many
commercial DNase I formulations are contaminated with residual RNases. This
RNase contamination can destroy or degrade valuable RNA samples prior to
reverse transcription. Laboratory comparisons have shown that Sigma′s
1 kit
Oligo(dT)23, Anchored
The Anchored Oligo(dT)23 Primers have 23 thymidine residues and one G, C, or
A residue (the anchor) at the 3′ end. This anchor ensures the oligo(dT)23 primer
binds at the beginning of the message such that there are no long regions of
unusable sequence. Anchored oligo(dT)23 primers may provide an advantage
over standard oligo(dT) primers when generating cDNA from poly(A)+ RNA.
 70 μM in H2O
0.1 mL sufficient for 100 RT-PCR reactions (as described in the Technical
Bulletin for Product Codes HSRT100 and HSRT20)
O4387-.1ML
0.1 mL
Random Nonamers
Random Nonamers are random sequences of nine deoxyribonucleotides
(9-mers). Random nonamers may be used as universal primers in first strand
cDNA synthesis, cDNA library construction, RT-PCR and other applications.
 50 μM in H2O
0.1 mL sufficient for 100 RT-PCR reactions (as described in the Technical
Bulletin for Product Codes HSRT100 and HSRT20.)
R7647-100UL
100 μL
Real-time PCR
Standard qPCR Reagents: SYBR® Green
Real-time PCR
Standard qPCR Reagents
10
SYBR® Green
SYBR Green JumpStart Taq ReadyMix combines the performance enhancements of JumpStart Taq antibody for hot start PCR with SYBR Green I and the
convenience of an easy-to-use ReadyMix solution. This ready-to-use mixture of
SYBR Green I, JumpStart Taq DNA polymerase, 99% pure deoxynucleotides and
reaction buffer is provided in a 2× concentrate for ease of use. Simply add
25 μL of the 2× mix to DNA template, primers and water. The JumpStart Taq
antibody inactivates the DNA polymerase at room temperature. When the
temperature is raised above 70 °C in the first denaturation step of the cycling
process, the complex dissociates and the polymerase becomes fully active.
JumpStart Taq DNA polymerase prevents non-specific amplification resulting
in more accurate CT values.
SYBR Green JumpStart Taq ReadyMix is recommended for single product realtime amplification experiments and can also be used for PCR optimization
prior to manufacture of fluorescent-labeled probes. Fluorescent labeled probes
are not recommended for use with SYBR Green I dye.
SYBR Green I, a commonly used fluorescent DNA binding dye, binds all
double-stranded DNA and detection is monitored by measuring the increase
in fluorescence throughout the cycle. SYBR Green I has an excitation and
emission maxima of 494 nm and 521 nm, respectively. The instrument settings
for ROX reference dye are satisfactory for the measurement of the Reference
Dye for Quantitative PCR. Specificity of Sigma′s SYBR based QPCR detection is
greatly enhanced by the incorporation of a hot-start mediated taq polymerase,
JumpStart Taq.
• Delivers the benefits of antibody-inactivated hot-start PCR with SYBR Green
detection in a ReadyMix ideal for high throughput applications; only primers
and template are required.
• JumpStart Taq DNA polymerase prevents amplification of non-specific
products, resulting in increased efficiency and higher target yield.
• SYBR Green JumpStart Taq ReadyMix for SYBR based QPCR is formulated
with MgCl2 or packaged with a separate vial for ease of optimization.
ReadyMixes are compatible with tube- and plate-based instruments.
ΔRn
SYBR® Green JumpStart™ Taq ReadyMix™
1
0.1
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40
Cycle
Ct values for the lambda amplicon using SYBR Green JumpStart Taq ReadyMix.
Quantitative PCR (qPCR) was performed on pBac-2cp. Initial template copy number was 106 and was
diluted 10-fold in subsequent wells. Threshold cycles (Ct) were determined using the ABI PRISM 7700
Sequence Detection software, and were found to be 15.304 (106), 18.848 (105), 22.883 (104), 26.208 (103),
29.821 (102), 33.398 (101), 37.038 (100), and 40 (0).
 for quantitative PCR, MgCI2 in buffer
S4438-20RXN
20 reactions
S4438-100RXN
100 reactions
S4438-500RXN
500 reactions
 for quantitative PCR, without MgCl2 in buffer
Magnesium chloride is provided separately for optimization.
S5193-20RXN
20 reactions
S5193-100RXN
100 reactions
S5193-400RXN
400 reactions
SYBR® Green JumpStart™ Taq ReadyMix™ for High Throughput
qPCR
SYBR Green ReadyMix for High Throughput Quantitative PCR combines the
performance enhancements of JumpStart Taq and SYBR Green I in an easy-touse ReadyMix solution that incorporates ROX dye for ABI and other real time
instrument applications. The ReadyMix includes a detection fluor, internal
standard and reagents for PCR making it the ideal solution for performing
high-throughput quantitative PCR.
• SYBR Green I dye binds to double-stranded DNA and is ideal for quantifying
any DNA sequence. Detection is monitored by measuring the increase in
fluorescence throughout cycling.
• JumpStart Taq DNA polymerase prevents amplification of non-specific
products while increasing target yield.
• Internal Reference Dye is provided for reaction normalization. Maximum
excitation and emission is 586 nm and 605 nm, respectively.
• SYBR Green JumpStart Taq ReadyMix reduces preparation time and the risk
of contamination from multiple pipetting steps.
17
sigma.com
 SYBR® Green qPCR reagent for Roche LightCycler® capillary systems
concentration .............................................................................. 1 units/reaction (20 μL reaction volume)
 SYBR® Green qPCR reagent, passive reference dye included
S9194-20RXN
20 reactions
S9194-400RXN
400 reactions
S9194-2000RXN
2000 reactions
Fluorescence (F1)
100.000
10.000
1.000
0.100
SYBR® Green JumpStart™ Taq ReadyMix™ for Quantitative PCR,
Capillary Formulation
SYBR Green JumpStart Taq ReadyMix, Capillary formulation combines the
advantages of a hot start enzyme, JumpStart Taq, in a 2× concentrate
ReadyMix specifically designed for use with capillary instruments, such as the
Roche LightCycler® real-time thermal cycler. SYBR Green JumpStart Taq
ReadyMix is an optimized formulation containing SYBR Green I dye, JumpStart
Taq DNA Polymerase, 99% pure deoxynucleotides, buffer and stabilizers.
SYBR Green Taq ReadyMix is recommended for single product real-time
amplification experiments and may also be used for evaluation of primer
sequences prior to manufacture of fluorescent-labeled probes. Fluorescent
labeled probes are not recommended for use with SYBR Green I dye.
SYBR Green I, a commonly used fluorescent DNA binding dye, binds all
double-stranded DNA and detection is monitored by measuring the increase
in fluorescence throughout the cycle. SYBR Green I has an excitation and
emission maxima of 494 nm and 521 nm, respectively. Specificity of Sigma′s
SYBR based QPCR detection is greatly enhanced by the incorporation of a hotstart mediated taq polymerase, JumpStart Taq.
The JumpStart Taq antibody inactivates the DNA polymerase at room
temperature. When the temperature is raised above 70 °C in the first
denaturation step of the cycling process, the complex dissociates and the
polymerase becomes fully active. JumpStart Taq DNA polymerase prevents
non-specific amplification resulting in more accurate CT values.
To prepare a reaction, 10 μL of ReadyMix is added to primers, template and
water for a final reaction volume of 20 μL.
• Convenient 2× concentrate ReadyMix specifically designed for use with
capillary instruments such as the Roche LightCycler and is ideal for high
throughput applications
• Increased specificity & target yield - JumpStart Taq polymerase prevents nonspecific product resulting in more accurate CT values and improved
quantitation
A tube of 25 mM MgCl2 is provided for easy optimization of the QPCR reaction.
18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45
Cycle Number
Quantitative PCR on human genomic DNA.
Human genomic DNA template was diluted 10-fold in subsequent wells; concentrations were 30 ng,
3 ng, 300 pg, 30 pg, 3 pg. Sigma′s SYBR Green Taq ReadyMix, Capillary Formulation was used with
specific primers to amplify a 250 bp PCR product of the β-actin gene.
S1816-20RXN
20 reactions
S1816-100RXN
100 reactions
S1816-400RXN
400 reactions
RT-qPCR
SYBR® Green Quantitative RT-qPCR Kit
The SYBR Green Quantitative RT-PCR kit provides a highly sensitive method for
the quantitative analysis of gene expression. Sigma′s QRT-PCR ReadyMix
combines the advantages of Moloney Murine Leukemia Virus Reverse
Transcriptase (M-MLV RT) and JumpStart Taq in a convenient ReadyMix.
1 kit sufficient for 100 reactions at 50 μL each
10.000 –
Fluorescence (F1)
18
1.000 –
0.100 –
0.010 – | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40
Cycle Number
One-step SYBR® Green RT-qPCR on a capillary-based system.
Assay for a ß actin 187 bp product was performed in duplicate on human total RNA from cell line HeLa
S3. Total RNA was diluted 10-fold in subsequent capillaries with concentrations ranging from 500 ng to
5 pg.
 One step SYBR® Green RT-qPCR with MMLV & hot-start Taq DNA
Polymerase
Components
SYBR® Green Taq ReadyMix™ for Quantitative RT-PCR 2 x 25 mL
Molony Murine Leukemia Virus Reverse Transcriptase (M-MLV™) 5000 units
10X PCR Buffer (Sigma P2192)
25 mM MgCl2 1.5 mL/vial
Reference Dye for Quantitative PCR (Sigma R4526) 0.3 ml/vial
QR0100-1KT
1 kit
Real-time PCR
Standard qPCR Reagents: Probe-based
B) 40
Probe-based
JumpStart Taq ReadyMix for Quantitative PCR combines the advantages of a
hot start enzyme with a ready-to-use mix for high throughput, quantitative
PCR (qPCR). It is formulated without a detection chemistry making it suitable
for use with a variety of formats including dual-labeled probes, molecular
beacons, or double stranded binding dyes such as SYBR Green I.
The ReadyMix contains JumpStart Taq DNA Polymerase, 99% pure deoxynucleotides, and buffer in an optimized 2× concentrate. To prepare a reaction,
25 μL of the ReadyMix is added to primers, template, detection chemistry, and
water for a total reaction volume of 50 μL. Set up can be performed at room
temperature since the JumpStart Taq antibody renders the Taq DNA
polymerase inactive. During the first denaturation cycle, the antibody
dissociates from the enzyme and full activity is restored. No special
preparations or protocol changes are required.
• Ultimate Convenience. ReadyMixes contain all components necessary for
QPCR, simply add fluorescent detection chemistry, primers, and template
• Greater Specificity & Increased Target Yield. JumpStart Taq prevents nonspecific amplification and increased target yield
• Maximum Flexibility. Suitable for use with a variety of detection chemistries
including molecular probes and double-stranded binding dyes such as SYBR
Green I
Sigma′s Reference Dye for Quantitative PCR is included separately with this
ReadyMix for normalization of the reaction data. The dye has a maximum
excitation of 586 nm, and a maximum emission of 605 nm. The instrument
settings for ROX reference dye are satisfactory for the measurement of the
Reference Dye for Quantitative PCR. A tube of 25 mM MgCl2 is also provided
for easy optimization of the qPCR reaction.
Rn
A)
Threshold Cycle (CT)
35
JumpStart™ Taq ReadyMix™ for Quantitative PCR
30
25
20
15
10
5
Supplier A
y = –3.5335x + 30.52
R2 = 0.9991
JumpStart Taq ReadyMix
y = –3.338x + 26.869
R2 = 0.9974
0
-2
-1
0
Log ng DNA template
1
Quantitative PCR was performed on human genomic DNA.
The template was diluted 10-fold in subsequent wells. A TaqMan® probe and primers specific for a
250 bp PCR product of the β-actin gene were used with Sigma′s JumpStart Taq ReadyMix for
Quantitative PCR (Cat. No. D7440) or a master mix from Competitor A. The JumpStart Taq ReadyMix (in
red) has better amplification efficiency (A), resulting in lower CT values than Competitor A (in blue) (B).
 For probe-based real-time PCR
Components
JumpStart™ Taq ReadyMix™ (Sigma P2893) 2x
25 mM MgCl2 1.5 μL
Reference Dye for Quantitative PCR (Sigma R4526) 100x
D7440-20RXN
20 reactions
D7440-100RXN
100 reactions
D7440-400RXN
400 reactions
JumpStart™ Taq ReadyMix™ for High Throughput
Quantitative PCR
JumpStart Taq ReadyMix For High Throughput Quantitative PCR combines the
performance enhancements of JumpStart Taq Antibody for hot start PCR with
the convenience of an easy-to-use reaction mixture that incorporates the
internal reference dye for ABI and other real time instrument applications. The
ReadyMix contains JumpStart Taq DNA polymerase, 99% pure deoxynucleotides and reaction buffer. It is provided in a 2× concentrate. Simply add an
equal volume of the 2× ReadyMix to a 2× mixture DNA template, primers and
fluorescent probe.
JumpStart Taq ReadyMix for High Throughput Quantitative PCR is a reaction
formulation used for high throughput hot-start quantitative PCR methods. It
contains the hot start DNA polymerase reaction mixture with ROX dye as an
internal reference for normalization of reactions.
5.000
4.500
4.000
3.500
3.000
2.500
2.000
1.500
1.000
0.500
0.000
-0.500
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 3032 34 36 38 40
Cycle
• To prepare a typical QPCR reaction, mix 25 μL of JumpStart ReadyMix Taq,
fluorescent probe, desired amount of magnesium chloride (above 1.5 mM
final concentration), template DNA, and primers in a final volume of 50 μL.
Reaction volumes can be scaled down, if desired.
• JumpStart Taq DNA polymerase prevents non-specific product formation,
and allows assembled PCR reactions to be placed at room temperature for
up to two hours without compromising the performance.
• Internal Reference Dye is provided for reaction normalization.
• When performing large numbers of PCR reactions, JumpStart ReadyMix Taq
can save a significant amount of preparation time, reduce the risk of
contamination from multiple pipetting steps, and provide consistent batchto-batch and reaction-to-reaction performance.
19
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sigma.com
Use 2.5 units per reaction.
Fast qPCR Reagents
SYBR® Green
LuminoCt® SYBR® Green qPCR ReadyMix™
 Ready-to-use 2x mix for qPCR with ROX
D6442-20RXN
20 reactions
D6442-400RXN
400 reactions
D6442-2000RXN
2000 reactions
RT-qPCR
Quantitative RT-PCR ReadyMix™
 One step RT-qPCR for probe-based methods, MMLV & hot-start Taq
The Quantitative RT-PCR ReadyMix kit includes Moloney Murine Leukemia Virus
Reverse Transcriptase (M-MLV RT), for efficient first strand cDNA. In addition,
the ReadyMix is blended with JumpStart Taq DNA polymerase, 99% pure
deoxynucleotides, buffer, glass passivator, and stabilizers, and is provided as a
2× concentrate for convenience. The kit provides high specificity, reduced risk
of contamination, and increased reproducibility.
1 kit sufficient for 100 reactions at 50 μL each
Components
• Kit is designed to perform SYBR Green-based qPCR assays on commonly
available real-time instrument platforms
• ReadyMix requires the addition of reference dye (provided in kit) when being
used on real-time instruments that require ROX passive reference dye for
normalization of qPCR assays
• Kit is not compatible with qPCR instruments that utilize glass capillary tubes
• Assay results in as little as 25 minutes
• Unsurpassed accuracy, precision, and sensitivity
• Virtually eliminates the need to optimize assay parameters
Probe Based qRT-PCR ReadyMix 2x
Moloney Murine Leukemia Viral Reverse Transcriptase (M-MLV RT) 5000 units
10X PCR Buffer (Sigma P2192) 1.5 mL / vial
Magnesium chloride (Sigma M8787) 25 mM
Reference Dye for Quantitative PCR (Sigma R4526) 100x
QR0200-1KT
LuminoCt SYBR Green qPCR ReadyMix is designed to deliver unsurpassed
assay speeds without sacrificing accuracy, precision, or sensitivity. Extensive
design and validation of the ReadyMix chemistry has virtually eliminated the
need for optimization when LuminoCt is used in conjunction with properly
designed primers. Inclusion of JumpStart™ Taq antibody in the ReadyMix
eliminates polymerase activity at ambient temperatures without causing the
performance issues associated with chemically modified hot-start Taq. This
allows for very rapid activation of the Taq and delivers unparalleled assay
sensitivity, while allowing for benchtop reaction setup.
1 kit
 For fast SYBR Green quantitative PCR
Components
LuminoCt SYBR Green qPCR ReadyMix 2x
100X ROX internal reference dye
L6544-100RXN
100 reactions
L6544-500RXN
500 reactions
L6544-2000RXN
2000 reactions
KiCqStart® SYBR® Green qPCR ReadyMix™
KiCqStart SYBR Green qPCR ReadyMix is a 2X concentrated, ready-to-use
reaction cocktail that contains all components, except primers and template,
for real-time quantitative PCR (qPCR) This unique combination of proprietary
buffer, stabilizers, and Hot-Start Taq DNA polymerase delivers maximum PCR
efficiency, sensitivity, specificity and robust fluorescent signal using fast, or
conventional, cycling protocols with SYBR Green qPCR.
Highly specific amplification is crucial to successful qPCR with SYBR Green I dye
technology because this dye binds to and detects any dsDNA generated
during amplification. Hot-Start Taq DNA polymerase is antibody mediated to
be inactive prior to the initial PCR denaturation step. The optimized
formulation includes SYBR Green I dye, an antibody-mediated Hot-Start Taq
DNA polymerase, dNPTs, MgCl2 and proprietary buffers and stabilizers.
Real-time PCR
Fast qPCR Reagents: SYBR® Green
 For Bio-Rad, Cepheid, Eppendorf, Illumina, Corbett, and Roche systems
• Highly efficient and sensitive real-time PCR results
• Little/no optimization required
KiCqStart SYBR Green qPCR ReadyMix is available for the following platforms:
Bio-Rad CFX384™, Bio-Rad CFX96™, Bio-Rad MiniOpticon™, Bio-Rad/MJ
Chromo4™, Bio-Rad/MJ Opticon™ 2, Bio-Rad/MJ Opticon™, Cepheid
SmartCycler®, Eppendorf Mastercycler® ep realplex, Eppendorf Mastercycler® ep
realplex2 s, Illumina Eco qPCR, Qiagen/Corbett Rotor-Gene® 3000,
Qiagen/Corbett Rotor-Gene® 6000, Qiagen/Corbett Rotor-Gene® Q, Roche
LightCycler® 480
KiCqStart™ SYBR® Green ReadyMix™
Competitor Q Fast SYBR® Green PCR Kit
36
Slope
R2
PCR Eff.
Cycle Threshold
34
32
1250 reactions
KCQS00-5000RXN
5000 reactions
KiCqStart SYBR Green qPCR ReadyMix. Low ROX is available for the following
platforms: Applied Biosystems 7500, Applied Biosystems 7500 Fast, Applied
Biosystems ViiA 7, Stratagene Mx3000P®, Stratagene Mx3005P®, Stratagene
Mx4000™
28
26
24
22
20
1
2
3
4
5
Log Initial Quantity (pg HeLa total RNA)
3000
KCQS01-250RXN
250 reactions
KCQS01-1250RXN
1250 reactions
KCQS01-5000RXN
5000 reactions
 with ROX™ for ABI instruments
KiCqStart SYBR Green qPCR ReadyMix. with ROX is available for the following
platforms: Applied Biosystems 5700, Applied Biosystems 7000, Applied
Biosystems 7300, Applied Biosystems 7700, Applied Biosystems 7900, Applied
Biosystems 7900 HT Fast, Applied Biosystems 7900 HT, Applied Biosystems
StepOnePlus, Applied Biosystems StepOne
2000
1000
0
0
5
10
15
20
25
30
35
40
PCR Cycle
Fast-cycling KiCqStart® SYBR® Green qPCR ReadyMix™ demonstrates equal performance to conventional
SYBR Green protocol and reagents.
Fast Cycling
Conventional Cycling
Cycle Threshold
1250 reactions
KCQS02-5000RXN
5000 reactions
 iQ™, with fluorescein for Bio-Rad systems
KCQS03-250RXN
250 reactions
KCQS03-1250RXN
1250 reactions
28
KCQS03-5000RXN
5000 reactions
26
24
20
1
2
3
4
5
Log Initial Quantity (pg HeLa total RNA)
3000
2000
1000
0
0
250 reactions
KCQS02-1250RXN
30
32
22
4000
KCQS02-250RXN
KiCqStart SYBR Green qPCR ReadyMix, iQ is available for the following
platforms: Biorad iCycler iQ™, BioRad iQ™5, BioRad MyiQ™
Slope -3.266 -3.217
-0.997 -0.998
R2
PCR Eff. 102.4% 104.6%
34
Relative Fluorescence
250 reactions
KCQS00-1250RXN
 Low ROX™, for ABI and Stratagene instruments
30
4000
Relative Fluorescence
-3.266 -3.305
-0.997 -0.996
102.4% 100.7%
KCQS00-250RXN
5
10
15
20
25
30
35
40
PCR Cycle
Target gene was amplified from log-fold dilutions of total HeLa cell cDNA (100 ng to 10 pg) using
KiCqStart® SYBR® Green qPCR ReadyMix™ or Competitor Q according to each manufacturer’s protocol.
Plots represent averages of quadruplicate reactions.
21
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sigma.com
MicroRNA RT-qPCR Assays
Probe-based
MystiCq® microRNA cDNA Synthesis Mix
LuminoCt® qPCR ReadyMix™
LuminoCt qPCR ReadyMix is designed to deliver unsurpassed assay speeds
without sacrificing accuracy, precision, or sensitivity. Extensive design and
validation of the ReadyMix chemistry has virtually eliminated the need for
optimization when LuminoCt is used in conjunction with properly designed
primers and probes. Inclusion of JumpStart™ Taq antibody in the ReadyMix
eliminates polymerase activity at ambient temperatures without causing the
performance issues associated with chemically modified hot-start Taq. This
allows for very rapid activation of the Taq and delivers unparalleled assay
sensitivity, while allowing for benchtop reaction setup.
• Kit is designed to perform probe-based qPCR assays on commonly available
real-time instrument platforms.
• ReadyMix requires the addition of reference dye (provided in kit) when being
used on real-time instruments that require ROX passive reference dye for
normalization of qPCR assays.
• Kit is not compatible with qPCR instruments that utilize glass capillary tubes.
• Unsurpassed accuracy, precision, and sensitivity
• Virtually eliminates the need to optimize assay parameters
LuminoCt qPCR ReadyMix, Catalog Number L5794, contains optimized
concentrations of Tris-HCl, pH 8.3, KCl, dNTPs (dATP, dCTP, dGTP, TTP),
stabilizers, MgCl2 and JumpStart Taq DNA Polymerase. Provided as 100, 500
and 2000 reactions (25 μL mix in a 50 μL reaction volume).
100x ROX internal reference dye, Catalog Number R4526. Optional, for use with
machines compatible with an internal reference dye, e.g., ABI and Stratagene.
Starting with total RNA or samples pre-enriched or microRNAs, the MystiCq
microRNA cDNA Synthesis Mix kit contains all the components necessary to
convert mature microRNAs into cDNA templates for qPCR. Since microRNAs
are not naturally polyadenylated, the first step in the cDNA synthesis process is
to polyadenylate the microRNAs through a poly(A) polymerase reaction. Next,
the ReadyScript reverse transcriptase, an oligo-dT adapter primer and other
required reagents are added to convert the polyadenylated microRNAs into
cDNA. The adapter primer includes a unique sequence at the 5’ end that is
complementary to the sequence of the MystiCq Universal PCR Primer, another
component of the MystiCq microRNA qPCR Assay System.The kit also contains
a Positive Control Primer that is specific to SNORD44, a small nucleolar RNA
that is widely expressed in most human tissues. Sufficient amounts of the Poly
(A) Tailing Buffer and the MicroRNA cDNA Reaction mix are included in the kit
to allow for the use of no poly(a) polymerase and no reverse transcriptase
control reactions. To analyze individual microRNAs, use the MystiCq Universal
PCR Primer (which targets the adapter primer sequence) together with the
MystiCq microRNA qPCR Assay Primer and the MystiCq microRNA SYBR Green
qPCR ReadyMix.
• Separate polyadenylation and reverse transcription steps allow for the
possibility of optimization of protocol if necessary
• cDNA produced may be archived for future use
• Single RT reaction can provide up to 1000 qPCR reactions
• Includes a positive control primer for use with most human tissues
Components
 For fast probe-based quantitative PCR
Poly (A) Tailing Buffer (5X)
Human Positive Control Primer
Nuclease-free Water
MystiCq® Universal PCR Primer
Poly (A) Polymerase
microRNA cDNA Reaction Mix
ReadyScript® Reverse Transcriptase
Components
MIRRT-25RXN
25 reactions
LuminoCt qPCR ReadyMix
100X ROX internal reference dye
MIRRT-100RXN
100 reactions
L6669-100RXN
100 reactions
L6669-500RXN
500 reactions
L6669-2000RXN
2000 reactions
Real-time PCR
MicroRNA RT-qPCR Assays
MystiCq® microRNA® SYBR® Green qPCR ReadyMix™
MystiCq® MicroRNA qPCR Primers
• Exclusively for use with the MystiCq microRNA qPCR Assay Primers, Control
Primers, and Universal PCR primer
• 2X concentrated for easy reaction setup with hot-start Taq, dNTPs and
optimized buffer
• Passive reference dye (if required) included at the optimal concentration
MystiCq microRNA qPCR Assay Primers
 Formulated for systems that do not require normalization
These Control Primers are an integral part of the MystiCq microRNA qPCR
Assay System. They have been designed to target specific small RNAs with the
MystiCq Universal PCR Primer and the MystiCq microRNA SYBR Green qPCR
ReadyMix on cDNA templates generated by the MystiCq microRNA cDNA
Synthesis Mix.
These Assay Primers are an integral part of the MystiCq microRNA qPCR Assay
System. They have been designed to target specific microRNAs with the
MystiCq Universal PCR Primer and the MystiCq microRNA SYBR Green qPCR
Synthesis Mix.
MystiCq microRNA qPCR Control Primers
MystiCq microRNA SYBR Green qPCR ReadyMix is compatible with the
following platforms: Bio-Rad CFX384™, Bio-Rad CFX96™, Bio-Rad MiniOpticon™,
Bio-Rad/MJ Chromo4™, Bio-Rad/MJ Opticon 2, Bio-Rad/MJ Opticon, Cepheid
SmartCycler®, Eppendorf Mastercycler® ep realplex, Eppendorf Mastercycler
ep realplex2 s, Illumina Eco qPCR, Qiagen/Corbett Rotor-Gene® 3000,
Qiagen/Corbett Rotor-Gene 6000, Qiagen/Corbett Rotor-Gene Q, Roche
LightCycler® 480.
MIRRM00-100RXN
100 reactions
MIRRM00-500RXN
500 reactions
MIRRM00-2000RXN
2000 reactions
 Low ROX™ formulation for miRNA RT-qPCR
MystiCq microRNA SYBR Green qPCR ReadyMix, Low ROX™, is compatible with
the following platforms: Applied Biosystems 7500, Applied Biosystems 7500
Fast, Applied Biosystems ViiA 7, Stratagene Mx3000P®, Stratagene Mx3005P®,
Stratagene Mx4000™.
MIRRM01-100RXN
100 reactions
MIRRM01-500RXN
500 reactions
MIRRM01-2000RXN
2000 reactions
 with ROX™, formulation for miRNA RT-qPCR
MystiCq microRNA SYBR Green qPCR ReadyMix, ROX™, is compatible with the
following platforms: Applied Biosystems 5700, Applied Biosystems 7000,
Applied Biosystems 7300, Applied Biosystems 7700, Applied Biosystems 7900,
Applied Biosystems 7900 HT Fast, Applied Biosystems 7900 HT, Applied
Biosystems StepOnePlus, Applied Biosystems StepOne.
MIRRM02-100RXN
100 reactions
MIRRM02-500RXN
500 reactions
MIRRM02-2000RXN
2000 reactions
 Formulation for miRNA RT-qPCR on iQ Bio-Rad platforms
MystiCq microRNA SYBR Green qPCR ReadyMix, iQ™, is compatible with the
following platforms: Biorad iCycler iQ, BioRad iQ5, BioRad MyiQ™.
MIRRM03-100RXN
100 reactions
MIRRM03-500RXN
500 reactions
MIRRM03-2000RXN
2000 reactions
MystiCq Universal PCR Primer
These PCR Primers are an integral part of the MystiCq microRNA qPCR Assay
System. It has been designed to work specifically with the MystiCq microRNA
qPCR Assay or Control Primers and the MystiCq microRNA SYBR Green qPCR
Synthesis Mix.
Product Description
MystiCq microRNA qPCR Assay Primer
MystiCq microRNA qPCR Control Primer
MystiCq Universal PCR Primer
Check sigma.com/mysticq for up-to-date product
Cat. No. Range
MIRAP00001-250RXN through MIRAP01434-250RXN
MIRCP00001-250RXN through MIRCP00016-250RXN
MIRUP-500RXN
listings.
Related Products for Real-time PCR
Reference Dye for Quantitative PCR
Sigma′s Reference Dye for qPCR is a proprietary dye for use with real-time PCR.
It is used for normalization of reaction data when using SYBR Green, molecular
probes, or dual-labeled probe chemistries for real-time detection. The
Reference Dye is supplied as a 100× solution with a maximum excitation and
emission of 586 nm and 605 nm, respectively. Instrument settings for ROX
reference dye are satisfactory for the measurement of Reference Dye for qPCR.
 100 ×
sufficient for ≥600 reactions
R4526-.3ML
0.3 mL
R4526-5ML
5 mL
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sigma.com
GenomePlex® Whole Genome Amplification (WGA) Kit
WGA is suitable for use with genomic DNA, whole blood, buffy coats, buccal
swabs, and cultured cells. Amplification requires only a small amount of
starting material (1 μL blood, single cheek swab, or 10 ng of gDNA), which
after PCR produces a yield of 5 - 10 μg per reaction. The amplified DNA can be
purified and stored or used for downstream analyses including ABI′s TaqMan®
assays, microsatellite analysis, or sequencing. GenomePlex gives a complete
representation of the entire genome with no detectable allele bias.
 Kit for whole genome amplification from a variety of DNA sources
including FFPE tissue
Components
10X Fragmentation Buffer 55 μL
1× Library Preparation Buffer 110 μL
Library Stabilization Solution 55 μL
Library Preparation Enzyme 55 μL
10X Amplification Master Mix 410 μL
Nuclease-Free Water 2 x 1.5 mL
Control Human Genomic DNA 10 μL
GenomePlex Whole Genome Amplification (WGA) Kit utilizes a proprietary
technology based on random fragmentation of genomic DNA and conversion
of the resulting small fragments to PCR-amplifiable library molecules flanked
by universal priming sites. WGA is achieved by PCR amplification of the library
molecules using universal oligonucleotide primers.
WGA1-50RXN
50 reactions
• Flexibility to study DNA from any source
• No detectable locus or allele bias
• Low background amplification of DNA from single chromosomes and cells
• Compatible with a variety of microarray, capillary, and homogenous
platforms for sequencing, genotyping, CGH, FISH, ChIP
• Increased sensitivity and accuracy for population studies, mutation discovery,
and pharmacogenomics.
GenomePlex Complete Whole Genome Amplification Kit utilizes a proprietary
GenomePlex® Complete Whole Genome Amplification (WGA)
Kit
GenomePlex WGA Overview
Fragmentation
Anneal primers
Extend
2nd priming event
OmniPlex library formation
PCR amplify
Amplified Genomic DNA
GenomePlex WGA was performed on genomic DNA isolated from HT29 colon carcinoma cells and
from a healthy human male.
2.5 μg of WGA product was labeled with Cy3™ or Cy5 dye using the Genomic DNA Labeling Kit PLUS
(Agilent). The entire labeled sample was loaded onto an Agilent Human Genome CGH Microarray 105A.
Specific activities were between 28 and 43 for all samples, and always within 50% of samples being
compared. The dye swaps (A & B) demonstrate that there was no bias in the DNA labeling and the
aberrations detected are consistent with the HT-29 karyotype.
M
1
2
3
4
5
6
7
8
9
10
11
12
13
14
GenomePlex® WGA Reamplification Kit
GenomePlex WGA Reamplification Kit utilizes a proprietary amplification
method that is based on random fragmentation of genomic DNA and
conversion of the resulting small fragments to PCR-amplifiable library
molecules flanked by universal priming sites. WGA is achieved by PCR
amplification of the library molecules using universal oligonucleotide primers.
WGA sample 1
WGA sample 2
Avg C(t) Value
28
27
26
25
24
23
0
WGA was performed on increasing concentrations of human genomic DNA.
Amplification product can be detected on an agarose gel with as little as 10 pg of input DNA. Optimal
performance is found with 1 to 10 ng of starting material. Increasing the amount of input DNA to 100 ng
is not recommended.
Lanes
M: DNA Marker
1 and 2. no template
3 and 4. 1 pg DNA
5 and 6. 10 pg DNA
7 and 8. 100 pg DNA
9 and 10. 1 ng DNA
11 and 12. 10 ng DNA
13 and 14. 100 ng DNA
 Optimized kit with enzyme for amplifying a variety of DNA including
FFPE tissue
• Successfully produces a negative control sample without product
• Flexibility to study DNA from any source
• No detectable locus or allele bias
• Compatible with a variety of microarray, capillary, and homogenous
platforms for sequencing, genotyping, CGH, FISH, ChIP
• Increased sensitivity and accuracy for population studies, mutation discovery,
and pharmacogenomics
Components
Library Stabilization Solution
Library Preparation Enzyme
WGA DNA Polymerase
Water (Sigma W4502)
WGA2-10RXN
10 reactions
WGA2-50RXN
50 reactions
WGA2-500RXN
500 reactions
1
2
3
4
5
6
Number of Reamplifications
qPCR Effects of Reamplification.
Two 10 ng samples of purified human genomic DNA were amplified with WGA2. Each WGA product was
then reamplified five successive times with WGA3. qPCR was performed on each sample using primer
pairs representative of eight different single copy loci and the average C(t) was determined. Genomic
representation was maintained after five successive reamplifications.
 Reamplification of WGA product with minimal bias
• Successfully produces a negative control sample without product
• Maintains representation of the entire genome through subsequent
reamplifications
• Preserve previous source material by amplifying nanogram amounts of
starting genomic DNA into microgram yields (on average up to 500-fold) in
less than three hours
Components
S A5606 10X Amplification Master Mix
WGA DNA Polymerase
Deoxynucleotide Mix, 10 mM (Sigma D7295) 0.2 mL
WGA3-50RXN
50 reactions
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GenomePlex® Single Cell Whole Genome Amplification Kit
 Amplify genome of a single cell
Components
GenomePlex Single Cell Whole Genome Amplification Kit utilizes a proprietary
NAT1-01 Single Cell WGA Samples – TET
6.0E+01
1x Single Cell Library Preparation Buffer
10x Single Cell Lysis & Fragmentation Buffer
Proteinase K (Sigma P4850)
10X Amplification Master Mix
WGA DNA Polymerase
Control Human Genomic DNA
Nuclease-Free Water
WGA4-10RXN
3.0E+01
0.0E+00
10 reactions
WGA4-50RXN
50 reactions
WGA4-500RXN
500 reactions
SeqPlex DNA Amplification Kit
40
50
60
70
80
90
56.0
NAT1-01 Control
DNA Samples – FAM
NAT1-01 Control
DNA Samples – TET
8.0E+01
2.0E-02
4.0E+01
0.0E+00
0.0E+00
-2.0E-02
This kit is an extension of the WGA product line and has been developed to
integrate into the Illumina® (GAIIx sequencer), SOLiD™ System (from Life
Technologies), Ion Torrent (from Life Technologies) and other next generation
sequencing workflows, as a DNA amplification technology.
-4.0E+02
40
50
60
59.7
70
80
90
40
50
60
56.6
70
80
90
U937 Leukemia Single Cells were amplified using the GenomePlex Single Cell WGA procedure.
Ten nanograms of amplified DNA was then utilized in a MGB Eclipse SNP quantitative PCR reaction on
the ABI 7700. Dissociation curves allow allele determination based upon the melting temperature (Tm).
FAM and TET fluorescent signals distinguish homozygous wild and heterozygous mutant types. The
WGA amplified single cell DNA was tested for the NAT1-01 SNP as well as MTHFR01 and IGF1R-03 (data
not shown). Unamplified control DNA was evaluated with each group to serve as a positive control. The
data indicates that all amplified single cells were the same allele for each SNP tested and were, therefore,
unchanged by WGA.
Human Control
Chromosome 3
The SeqPlex DNA Amplification Kit for whole genome amplification (WGA) is
designed to facilitate next-generation sequencing (NGS) from extremely small
quantities or from degraded/highly fragmented DNA. The yields from
chromatin immunoprecipitation (ChIP) or formalin-fixed paraffin-embedded
tissue samples (FFPE) are often less than required for successful NGS library
preparation. The SeqPlex kit allows the user to pre-amplify these and other
small quantity/highly fragmented DNA samples for input into a NGS workflow.
Human Kidney Tumor
Chromosome 3
• Compatible with next generation sequencing platforms
• Patent pending random priming technology amplifies fragmented or intact
DNA (obtained by ChIP or any other protocol)
• Provides highly uniform amplification across the entire genome with
minimal sequence bias
• Directly compatible with validated sequencing library preparation protocols
on existing NGS platforms
• Enhanced primer design for more complete genomic coverage
 For use with high throughput sequencing technologies, Whole Genome
Amplification kit designed to facilitate Next Gen Sequencing
Components
10X Fragmentation Buffer
Library Preparation Buffer for SeqPlex
DNA Polymerase for SeqPlex
10X Primer Removal Buffer for SeqPlex
Primer Removal Enzyme for SeqPlex
Water (Sigma W4502)
SEQX-10RXN
DNA from normal and tumorigenic human kidney cells were amplified using the GenomePlex
Single Cell WGA Kit.
Amplified material was hybridized to metaphase BAC arrays to determine chromosomal karyotype. Data
that falls outside of the red line indicates chromsomal loss, while data that continues past the green line
suggests chromosomal amplification. As expected the control sample demonstrated a balanced
chromosomal copy number. Chromosome 3 for the amplified kidney tumor single cell displayed under
representation as depicted by the red bar. These results match previous microarray work using an
amplification method that took three days. GenomePlex technology accurately amplifies genomic
material down to single cell resolution.
Data is courtesy of Dr. Michael Speicher from Institute Human Genetics, TU Munich.
10 reactions
SEQX-50RXN
50 reactions
SEQX-500RXN
500 reactions
 For amplification of total RNA in less than 4 hours without 3′ bias
TransPlex® Whole Transcriptome Amplification Kit
Components
The WTA process involves two steps. In the first step, sample RNA is reverse
transcribed with non-self-complementary primers composed of a quasirandom 3′ end and a universal 5′ end. As polymerization proceeds, displaced
single strands serve as new templates for primer annealing and extension. The
resultant OmniPlex cDNA library, comprised of random, overlapping 100 - 1000
base fragments flanked by universal end sequence, is then amplified by PCR
with the universal primer to produce WTA product.
TransPlex, a Whole Transcriptome Amplification (WTA) method, allows for
representative amplification of nanogram quantities of total RNA in less than
4 hours without 3′-bias. Microgram quantities of amplification product
generated from tissue, cultured cells, formalin-fixed samples, or serum are
suitable for downstream applications such as qPCR and microarray analyses.
WTA Procedure
Input RNA
Add primers
First Strand cDNA
Second Strand cDNA
Unampliﬁed Total RNA
Normalized Δ C(t)
The WTA2 process involves two steps. In the first step, sample RNA is reverse
transcribed with non-self-complementary primers composed of a quasirandom 3′ end and a universal 5′ end. During this process, displaced single
strands serve as new templates for primer annealing and extension. The
resultant OmniPlex® cDNA library, comprised of random, overlapping 100 1000 base fragments flanked by universal end sequence. The 2nd step
amplifies the OmniPlex cDNA library by PCR using WTA2 polymerase and a
universal end primer to produce WTA2 product.
Components
Amplified cDNA
WTA dNTP Mix
Nuclease-Free Water
Library Synthesis Enzyme
Library Synthesis Solution
Amplification Mix
Library Synthesis Buffer
QPCR or microarray analysis
C
TransPlex® Complete Whole Transcriptome Amplification Kit
OmniPlex Library
PCR
B
50 reactions
 Complete Kit with optimized enzyme to amplify total RNA in <4 hours,
no 3′ bias
Denature
A
WTA1-50RXN
WTA2 is optimized to amplify RNA from formalin-fixed, paraffin-embedded
(FFPE) and other damaged or degraded samples. TransPlex, a Whole
Transcriptome Amplification (WTA) technology, allows for representative
amplification of low nanogram quantities of total RNA in less than 4 hours
without 3′-bias. Amplification products are suitable for applications such as
qPCR, micro array analysis, and cloning. The WTA2 kit contains the polymerase
needed to amplify the OmniPlex cDNA library.
Anneal and extend
3
2
1
0
-1
-2
-3
-4
-5
WTA Library Synthesis Buffer
WTA Library Stabilization Solution
WTA Library Synthesis Enzyme
WTA Amplification Master Mix
Water (Sigma W4502)
Deoxynucleotide Mix, 10 mM (Sigma D7295)
Ampliﬁed Total RNA
D
E
F
G
qPCR Primer Set.
Differential expression of seven mRNAs (cDNAs) in human liver and UHR (Universal Human Reference)
was measured by quantitative PCR. DC(t) values (UHR - Liver) were calculated for unamplified cDNA,
prepared directly from RNA by reverse transcription, and amplification product prepared from
25 nanograms of total RNA with the Transplex WTA Kit. (Normalization of C(t)s for liver or brain was
accomplished by subtracting the average DC(t) for all primer sets for a given tissue from each tissuespecific DC(t).) Uniformity between unamplified and amplified DC(t)s for individual mRNAs demonstrates
maintenance of differential patterns of RNA expression in liver and UHR when using the Transplex WTA
Kit. Universal Human Reference Total RNA is a product of Stratagene.
WTA2-10RXN
10 reactions
WTA2-50RXN
50 reactions
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Extract-N-Amp™ Kits
Blood Kits
REDExtract-N-Amp™ Blood PCR Kit
The REDExtract-N-Amp Blood PCR Kits contain all of the reagents necessary to
rapidly extract host genomic DNA from whole blood and amplify targets of
interest by PCR. This novel extraction system eliminates the need for any type
of purification, organic extraction, centrifugation, heating, filtration, or alcohol
precipitation. The kit also includes a PCR ReadyMix™, that uses an antibody
based hot start and is specially formulated for amplification directly from the
extract. The REDExtract-N-Amp Blood PCR Kit contains a tracking dye that
allows for convenient direct loading of PCR reactions onto agarose gels for
analysis.
For fresh or frozen blood samples, monolayers
Extract-N-Amp Blood PCR Kit
Remove aliquot of blood.
Genomic DNA is extracted from 10 μL of whole blood by simply adding the
Extraction Solution and incubating for 5 minutes at room temperature. The
Neutralization Solution is added to the extract to counteract inhibitory
substances prior to PCR. A portion of the DNA extract is then added to the
specially formulated PCR Mix.
• Efficient 8 minute prep allows greater speed and throughput
• No need for organic extraction, centrifugation or alcohol precipitation
• Hot start antibody for highly specific PCR amplification of genomic DNA
• Can be used with whole blood or blood cards
• Extract stable at 4 °C for at least 6 months
Mix blood with Lysis Solution.
Incubate room temp. 5 min.
8 minutes
Add Neutralization Solution.
Add aliquot of extract &
Mix aliquot with
REDExtract-N-Amp™
PCR Reaction Mix and primers.
Transfer to thermal cycler.
Directly load amplified PCR
product on gel.
Direct sequence from PCR products generated using the Extract-N-Amp™ Blood Kit.
A 547 bp product for human surfactant protein B was generated using the Extract-N-Amp Blood PCR kit.
The product was sequenced directly using BigDye® terminator chemistry. Sequencing reactions were
resolved on an ABI 3100.
Note: Some PCR products require further clean-up prior to sequencing. The GenElute™ PCR Clean-Up Kit
(Cat. No. NA1020) is recommended.
Frozen (-20 ˚C)
7 Day Old
Fresh
Frozen (-20 ˚C)
7 Day Old
Fresh
Frozen (-20 ˚C)
7 Day Old
Fresh
Frozen (-20 ˚C)
7 Day Old
M
Fresh
Blood Kits
 sufficient for 1000 extractions, sufficient for 1000 amplifications
XNABR-1KT
M
XNABRE-1KT
2 kb
1 kit
1 kit
1.5 kb
Extract-N-Amp™ Blood PCR Kit
1 kb
750 bp
The Extract-N-Amp Blood PCR Kits contain all of the reagents necessary to
rapidly extract host genomic DNA from whole blood and amplify targets of
interest by PCR. This novel extraction system eliminates the need for any type
of purification, organic extraction, centrifugation, heating, filtration or alcohol
precipitation. The kit also includes a PCR ReadyMix™, that uses an antibody
based hot start and is specially formulated for amplification directly from the
extract.
500 bp
300 bp
150 bp
50 bp
PCR analysis of genomic DNA isolated from blood using Sigma′s Extract-N-Amp™ Blood PCR Kit.
Extract-N-Amp™ Blood PCR Kit used to isolate genomic DNA from fresh, 7 day old, and frozen blood.
DNA was extracted and neutralized from 10 μL of whole blood in 5 minutes at room temperature using
the REDExtract-N-Amp™ Blood PCR kit. The PCR products were then amplified using the specially
formulated Hot Start PCR mix included in the kit. PCR products generated are 1.8 kb for carnitine
palmitoyltransferase II, 1.3 kb for a mitochondrial DNA control region, 547 bp for human surfactant
protein B, and 320 bp for the 5′ untranslated region of human major histocompatibility complex class II.
t=0
3 wk, 37 °C
1 wk, 37 °C
6 wk, 37 °C
6 mo, 37 °C
3.0
• Efficient 8 minute prep allows greater speed and throughput
• No need for organic extraction, centrifugation or alcohol precipitation
• Hot start antibody for highly specific PCR amplification of genomic DNA
• Can be used with whole blood or blood cards
2.5
XNAB2-1KT
2.0
ng DNA
Genomic DNA is extracted from 10 μL of whole blood by simply adding the
Extraction Solution and incubating for 5 minutes at room temperature. The
Neutralization Solution is added to the extract to counteract inhibitory
substances prior to PCR. A portion of the DNA extract is then added to the
specially formulated PCR Mix.
1 kit
1.5
XNAB2R-1KT
1 kit
1.0
0.5
XNAB2RE-1KT
0.0
1
2
3
Extract-N-Amp™ PCR ReadyMix™ for Blood
4
Extract #
Stability of Extract-N-Amp™ Blood Extracts.
Blood was drawn from 2 human volunteers into vacutainer tubes containing EDTA. Extractions were
performed in duplicate providing 4 samples total. Half the extracts were stored at 4 °C (recommended
storage conditions) and the other half at 37 °C (accelerated storage). Samples were removed at various
time intervals for testing. Stability was determined by monitoring yield from quantitative PCR using an
ABI 7700 instrument. The DNA standards used for the quantitative PCR were generated from the same
blood draw as the test samples, purified using the GenElute Blood Genomic DNA Kit (Cat. No. NA2000)
and stored as single aliquots at -20 °C. The PCR products were generated using primers for a 547 bp
product from human surfactant protein B (SPB) [Lin, Z. and Floros, J., BioTechniques, 29(3), 460 (2000)].
The results clearly show no loss of amplification of the SPB PCR product even after storage at 37 °C for
6 months. Similar results were obtained with storage at 4 °C.
XNABS-1KT
1 kit
XNAB-1KT
1 kit
XNABE-1KT
1 kit
1 kit
PCR ReadyMix™ is intended for use with Sigma′s Extract-N-Amp™ Blood PCR
Kits. All Extract-N-Amp Kits include a volume of PCR ReadyMix sufficient for at
least one PCR reaction per extraction. However, where additional amplifications are required, supplemental PCR ReadyMix may be needed.
 12 mL sufficient for 1000 amplifications
P8115-12ML
12 mL
REDExtract-N-Amp™ PCR ReadyMix™ for Blood
PCR ReadyMix is intended for use with Sigma′s Extract-N-Amp Blood PCR Kits.
All Extract-N-Amp Kit include a volume of PCR ReadyMix sufficient for at least
one PCR reaction per extraction. However, where additional amplifications are
required, supplemental PCR ReadyMix may be needed. The REDExtract-N-Amp
PCR ReadyMix contains a inert tracking dye that allows for convenient direct
loading of PCR reactions onto agarose gels for analysis.
P8240-12ML
12 mL
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sigma.com
Plant
For Plant Leaf Material
Extract-N-Amp Plant PCR Kit
REDExtract-N-Amp™ Plant PCR Kit
The REDExtract-N-Amp Plant PCR Kits contain all the reagents necessary to
rapidly extract genomic DNA from plant leaves and amplify targets of interest
by PCR. A novel Extraction Solution eliminates the need for conventional
freezing of plant tissues with liquid nitrogen, mechanical disruption, organic
extraction, column purification, or precipitation of DNA. The kit also includes a
PCR ReadyMix™, especially formulated for amplification directly from extract.
The REDExtract-N-Amp PCR ReadyMix contains a dye that acts as a tracking
dye and allows for convenient direct loading of PCR reactions onto agarose
gels for analysis.
Genomic DNA is extracted from 0.5 to 0.7 cm plant leaf disks that have been
cut with a standard paper punch and simply incubated in Extraction Solution
at 95 oC for 10 minutes. An equal volume of Dilution Solution is added to the
extract to neutralize inhibitory substances prior to PCR. A portion of the DNA
extract is then added to a PCR reaction containing primers and the REDExtractN-Amp ReadyMix, included in the kit.
• Genomic DNA for PCR in less than 15 minutes
• A PCR ReadyMix™, specially formulated for amplification directly from extract
• Hot Start antibody for highly specific PCR amplification of genomic DNA
Kits contain Dilution solution, Extraction solution, and REDExtract-N-Amp PCR
Reaction Mix.
Punch leaf disk.
Incubate with Extraction Solution.
Heat 95 °C for 10 min.
15 minutes
Add Dilution Solution.
Mix aliquot of extract with
REDExtract-N-Amp™
PCR ReadyMix and primers.
product on gel.
r
ol
ina tiva
pe
ntr
e
ree suckl aster enjam is sa e co
C
y
ia
b
tiv
ab
ne
gin Hone Coto Ficus Cann nega
Vir
Internal positive
control for
plant chloroplast
Cannabisspecific
product
PCR analysis of genomic DNA isolated from plant using Sigma′s Extract-N-Amp™ Plant PCR Kit.
The Extract-N-Amp™ Plant PCR Kit was used to extract and amplify genomic DNA from 0.5 cm leaf disks
from various plant sources. The products were generated from a 30-cycle duplex reaction containing
primers specific to plant chloroplast (upper band) and primers specific to a Cannabis sativa gene (lower
band). MW ladder is 100, 200, 400, and 800 bp. Data provided by Andy Hopwood, Forensic Science
Service, Birmingham, England.
Plant
t=0
3 wk, 37 °C
1 wk, 37 °C
6 wk, 37 °C
6 mo, 37 °C
1.5
• Single-step extraction of plant genomic DNA for PCR in less than 15 minutes
• A PCR ReadyMix™, specially formulated for amplification directly from extract
1.0
Kits contain Dilution solution, Extraction solution, and Extract-N-Amp PCR
Reaction Mix.
ng DNA
2.0
0.5
XNAP2-1KT
1 kit
0.0
1
3
2
4
XNAP2E-1KT
1 kit
Extract #
Stability of Extract-N-Amp™ Plant extracts.
Eight disks were punched from a corn leaf, and DNA was extracted with the Extract-N-Amp™ Plant PCR
Kit. Following extraction, two 4-μL aliquots from each sample were analyzed immediately by quantitative
PCR with SYBR® Green detection on an ABI PRISM® 7700. DNA standards for quantitative PCR were
purified DNA prepared from corn leaf tissue with the GenElute™ Plant Genomic DNA Miniprep Kit (Cat.
No. G2N70). Half of the leaf extracts were stored at 4 °C (recommended storage conditions) and the
other half at 37 °C (accelerated storage conditions). Quantitative PCR was repeated after 1, 3, and
6 months from extract at 4 °C, and after 1 week, 3 weeks, 6 weeks, and 6 months from extracts at 37 °C.
Results for storage at 37 °C are shown. Results indicate that extract will be stable 6 months and probably
several years at 4 °C. (data not shown)
XNAPS-1KT
1 kit
XNAP-1KT
1 kit
Extract-N-Amp™ PCR ReadyMix™
PCR ReadyMix is intended for use with Sigma′s Extract-N-Amp Plant PCR kit
and Extract-N-Amp Tissue PCR Kit. All Extract-N-Amp kits include a PCR
ReadyMix sufficient for one PCR reaction per extraction. However, if additional
PCR reactions are required, supplemental PCR ReadyMix may be needed.
 Amplifications to support Extract-N-Amp Plant and Extract-N-Amp Tissue
12 mL sufficient for 1000 amplifications
1 kit
XNAPR-1KT
XNAR-1KT
1.2 mL sufficient for 100 amplifications
1 kit
XNAPE-1KT
E3004-1.2ML
1.2 mL
E3004-12ML
12 mL
E3004-125ML
125 mL
1 kit
REDExtract-N-Amp™ PCR ReadyMix™
Extract-N-Amp™ Plant PCR Kit
The Extract-N-Amp Plant PCR Kits contain all the reagents necessary to rapidly
extract genomic DNA from plant leaves and amplify targets of interest by PCR.
A novel Extraction Solution eliminates the need for conventional freezing of
plant tissues with liquid nitrogen, mechanical disruption, organic extraction,
column purification, or precipitation of DNA. The kit also includes a PCR
ReadyMix™, especially formulated for amplification directly from extract.
Genomic DNA is extracted from 0.5 to 0.7 cm plant leaf disks that have been
cut with a standard paper punch and simply incubated in Extraction Solution
at 95 oC for 10 minutes. An equal volume of Dilution Solution is added to the
extract to neutralize inhibitory substances prior to PCR. A portion of the DNA
extract is then added to a PCR reaction containing primers and the Extract-NAmp PCR ReadyMix, included in the kit.
ReadyMix sufficient for one PCR reaction per extraction. However, where
additional PCR reactions are required, supplemental PCR ReadyMix may be
needed. The REDExtract-N-Amp PCR ReadyMix contains a inert tracking dye
that allows for convenient direct loading of PCR reactions onto agarose gels
for analysis.
R4775-1.2ML
1.2 mL
R4775-12ML
12 mL
R4775-125ML
125 mL
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sigma.com
For mouse tails, hair, animal tissue, buccal swabs
Tissue
Extract-N-Amp Tissue PCR Kit
REDExtract-N-Amp™ Tissue PCR Kit
The REDExtract-N-Amp Tissue PCR Kits provide all the reagents necessary to
rapidly extract DNA from a wide variety of cells and tissues and amplify targets
of interest by PCR. A novel extraction method eliminates the need for long
enzymatic digestions or homogenization. The kit also includes a specially
formulated hot start PCR reaction mix for amplification directly from the
extract. The REDExtract-N-Amp PCR ReadyMix contains an inert dye that acts as
a tracking dye and allows for convenient lading of PCR reactions onto agarose
gels for analysis.
Add mouse tail, buccal swab, etc. to Extraction
& Tissue Preparation Solution mixture
Incubate at room temp for 10 min.
The Extract-N-Amp Tissue PCR Kits provide all the reagents necessary to rapidly
extract DNA from a wide variety of cells and tissues and amplify targets of
interest by PCR. A novel extraction method eliminates the need for long
formulated hot start PCR ReadyMix™ for amplification directly from the extract.
The REDExtract-N-Amp PCR ReadyMix contains an inert dye that acts as a
tracking dye and allows for convenient loading of PCR reactions onto agarose
gels for analysis.
The kit contains validated protocols to extract and amplify genomic DNA from
mouse-tails, hair, animal tissue, saliva, and buccal swabs. In a typical procedure,
genomic DNA is extracted from a sample that has been incubated in the
Tissue Preparation Solution and Extraction Solution for 10 minutes at room
temperature. The sample is heated to 95 °C for 3 minutes and then mixed with
a third solution to neutralize inhibitory substances prior to PCR. A portion of
the DNA extract is then added to a PCR reaction containing primers and
REDExtract-N-Amp PCR ReadyMix, included in the kit.
• Perfect for quick genomic DNA isolation for genotyping
• Single-step extraction of genomic DNA for PCR in 15 minutes
• PCR ReadyMix, specially formulated for amplification directly from extract
• Extract stable at 4 oC for at least 6 months
Heat at 95 °C for 3 min.
15 minutes
Add Neutralization B Solution
Add aliquot of extract &
Mix aliquot with
REDExtract-N-Amp™
PCR Reaction Mix and primers.
product on gel.
Kits contain Tissue Preparation Solution, Extraction Solution, and REDExtract-NAmp PCR Reaction Mix.
PCR analysis of genomic DNA extracted from various samples using Sigma’s Extract-N-Amp™
Tissue PCR Kit.
The Extract-N-Amp Tissue PCR kit was used to extract and amplify genomic DNA from various sources.
Genomic DNA was extracted from the samples using the appropriate protocol for each sample type as
described in the Extract-N-Amp Tissue Technical Bulletin. In all cases, the extraction procedure was
completed in less then 15 minutes. The extracted DNA was then amplified using the specially
formulated hot start PCR mix, provided in the kit. The products generated are 1181 bp for the Interleukin
1 Beta gene in mouse and 1820 bp for the Carnitine palmitoyltransferase II in human. Markers are both
PCR Marker (Cat. No. P9577).
Tissue
ng DNA
t=0
3 wk, 37 °C
6 wk, 37 °C
2 mo, 37 °C
20
18
16
14
12
10
8
6
4
2
0
XNAT2R-1KT
1 kit
1
2
Extract #
Stability of DNA in mouse-tail extracts.
Mouse-tail samples were extracted according to the procedure in the Technical Bulletin for the ExtractN-Amp™ Tissue PCR Kit. The remaining mouse-tail tissue was removed from the samples for storage. 4 μL
aliquots were analyzed immediately by quantitative PCR with SYBR® Green detection on an ABI Prism
7700. DNA standards for quantitative PCR were purified DNA prepared from mouse tails using the
GenElute Mammalian Genomic DNA kit (Cat. No. G1N70) and stored as single use aliquots at –20 °C. The
mouse-tail extracts were stored at 37 °C (accelerated storage). Quantitative PCR was repeated after
3 weeks, 5 weeks and 2 months from extracts at 37 °C. Results for storage at 37 °C are shown. These
results suggest that extracts will be stable for at least 6 months at the recommended storage
temperature of 4 °C.
XNATS-1KT
1 kit
XNAT-1KT
1 kit
XNATR-1KT
1 kit
E3004-1.2ML
1.2 mL
E3004-12ML
12 mL
E3004-125ML
125 mL
for analysis.
Extract-N-Amp™ Tissue PCR Kit
The Extract-N-Amp Tissue PCR Kits provide all the reagents necessary to rapidly
extract DNA from a wide variety of cells and tissues and amplify targets of
formulated hot start PCR ReadyMix™ for amplification directly from the extract.
The kit contains validated protocols to extract and amplify genomic DNA from
mouse-tails, hair, animal tissue, saliva, and buccal swabs. In a typical procedure,
genomic DNA is extracted from a sample that has been incubated in the
Tissue Preparation Solution and Extraction Solution for 10 minutes at room
temperature. The sample is heated to 95 °C for 3 minutes and then mixed with
a third solution to neutralize inhibitory substances prior to PCR. A portion of
the DNA extract is then added to a PCR reaction containing primers and
Extract-N-Amp PCR ReadyMix, included in the kit.
• Perfect for quick genomic DNA isolation for genotyping
• Single-step extraction of genomic DNA for PCR in 15 minutes
• PCR ReadyMix is specially formulated for amplification directly from extract
• Extract stable at 4 oC for at least 6 months
Kits contain Tissue Preparation Solution, Extraction Solution, and Extract-NAmp PCR Reaction Mix.
XNAT2-1KT
1 kit
R4775-1.2ML
1.2 mL
R4775-12ML
12 mL
R4775-125ML
125 mL
Seed
REDExtract-N-Amp™ Seed PCR Kit
The REDExtract-N-Amp Seed PCR Kits provide all the reagents necessary to
rapidly extract DNA from a wide variety of plant seeds and amplify targets of
enzymatic digestions. The kit also includes a specially formulated hot start PCR
reaction mix for amplification directly from the extract. The REDExtract-N-Amp
PCR ReadyMix contains an inert dye that acts as a tracking dye and allows for
convenient loading of PCR reactions onto agarose gels for analysis.
The kit comes with validated protocols to extract and amplify genomic DNA
from corn, wheat, sunflower, cotton, soybean, sorghum, canola, and
Arabidopsis seeds. Additional protocols are available. In a typical procedure,
genomic DNA is extracted from a sample that has been ground and incubated
in the Seed Preparation Solution and Extraction Solution for 10 minutes at
room temperature. The sample is heated to 95 °C for 3 minutes and then
mixed with a third solution to neutralize inhibitory substances prior to PCR. A
portion of the DNA extract is then added to a PCR reaction containing primers
and the REDExtract-N-Amp PCR ReadyMix, included in the kit.
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• Perfect for quick genotyping results
• Rapid extraction of genomic DNA for PCR in 15 minutes
• No long enzymatic digestions
DNA extraction from wheat and subsequent PCR were performed using Sigma′s Extract-N-Amp
Seed PCR kit.
The PCR product of interest was purified with the GenElute™ PCR Clean-Up Kit (Cat. No. NA1020). The
chromatogram shows a portion of the sequence determination for the 964 bp acetyl-coenzyme A
carboxylase wheat PCR product. The sequence was obtained using the ABI BigDye® Terminator
Chemistry and the same primers as for the original PCR.
For Plant Seeds
Extract-N-Amp™ Seed PCR Kit
The Extract-N-Amp Seed PCR Kit provides all reagents necessary to rapidly
extract DNA from a wide variety of plant seeds and amplify targets of interest
by PCR. A novel extraction method eliminates the need for long enzymatic
digestions. The kit also includes a specially formulated hot start PCR reaction
mix for amplification directly from the extract.
The kit comes with validated protocols to extract and amplify genomic DNA
from corn, wheat, sunflower, cotton, soybean, sorghum, canola, and
Arabidopsis seeds. Additional protocols are available. In a typical procedure,
genomic DNA is extracted from a sample that has been ground and incubated
in the Seed Preparation Solution and Extraction Solution for 10 minutes at
room temperature. The sample is heated to 95 °C for 3 minutes and then
mixed with a third solution to neutralize inhibitory substances prior to PCR. A
portion of the DNA extract is then added to a PCR reaction containing primers
and the Extract-N-Amp PCR ReadyMix, included in the kit.
• Perfect for quick genotyping results
• Rapid extraction of genomic DNA for PCR in 15 minutes
• No long enzymatic digestions
Extract-N-Amp Seed PCR Kit
For Plant Seeds
Add ground seed to Extraction
& Seed Preparation Solution mixture.
Extract-N-Amp Seed PCR Kit
Incubate at 55 °C for 10 min.
Heat to 95 °C for 3 min.
Add ground seed to Extraction
& Seed Preparation Solution mixture.
15 minutes
Add Neutralization B Solution.
Incubate at 55 °C for 10 min.
Heat to 95 °C for 3 min.
15 minutes
REDExtract-N-Amp™
Add Neutralization B Solution.
REDExtract-N-Amp™
product on gel.
XNASS-1KT
product on gel.
1 kit
XNAS-1KT
1 kit
Seed
7
7
95
P
sis
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m
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ve
ea rn
on rghu nola abid heat gati 577
t
t
P9
Ne
W
Ar
Ca
Co
So
Co
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oy
S
SYBR® Green Tissue
SYBR® Green Extract-N-Amp™ Tissue PCR Kit
Wheat 964
Universal Plant
~400-700 bp
Genomic DNA was extracted from seeds using the protocol as described in the Extract-N-Amp™ Seed
Technical Bulletin. All extracts were then amplified using the specially formulated JumpStart™ PCR
ReadyMix™ and PCR primers multiplexed for both a universal chloroplast gene (~400-700) and the
acetyl-coenzyme A carboxylase gene specific to wheat (964 bp).
XNAS2-1KT
1 kit
The SYBR Green Extract-N-Amp Tissue PCR Kit contains all the reagents needed
for rapid extraction, amplification and detection of genomic DNA from mouse
tails and other animal tissues, buccal swabs, hair shafts, and saliva.
DNA is rapidly extracted from a tissue by incubating the sample with a mixture
of the Extraction Solution and the Tissue Preparation Solution at room
temperature for 10 minutes. After a 3-minute heat denaturing step, an equal
volume of Neutralization Solution B is added to the extract to neutralize
inhibitory substances and the extract is ready for real-time PCR in any platebased real-time thermal cycling system. An aliquot of the neutralized extract is
then combined with the Extract-N-Amp SYBR Green PCR ReadyMix™ and userprovided PCR primers.
Kit contains Extraction Solution, Neutralization Solution, Tissue Preparation
Solution, and Extract-N-Amp SYBR Green PCR ReadyMix™. The Extract-N-Amp
SYBR Green PCR ReadyMix is a 2X reaction mix containing SYBR Green, buffer,
salts, dNTPs, Taq polymerase and JumpStart™ Taq antibody. It is optimized
specifically for use with the extraction reagents and contains JumpStart Taq
antibody for hot start PCR to enhance specificity and SYBR Green I to act as a
nonspecific reporter for real-time PCR.
Rat Liver
Mouse Pancreas
Mouse Tail
No Template Control
40,000
E3004-1.2ML
1.2 mL
E3004-12ML
12 mL
E3004-125ML
125 mL
Fluorescence (dR)
30,000
20,000
10,000
0
2
4
6
8
10
12
14
16
18
20
22
24
26
28
30
32
34
36
38
40
Cycles
for analysis.
R4775-1.2ML
Human Buccal Swab
Human Hair
Human Saliva
1.2 mL
R4775-12ML
12 mL
R4775-125ML
125 mL
DNA extraction and amplification from various sources
Quantitative PCR was performed on DNA extracted from various starting materials using the SYBR®
Green Extract-N-Amp™ Tissue PCR Kit as outlined in the technical bulletin. As depicted above, the
Extract-N-Amp system can be used to quantify DNA from a wide range of sources.
XNATG-1KT
1 kit
XNATRG-1KT
1 kit
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sigma.com
SYBR® Green Extract-N-Amp™ Plant PCR Kit
SYBR® Green Extract-N-Amp™ PCR ReadyMix™
The SYBR Green Extract-N-Amp Plant PCR Kit contains all the reagents needed
for rapid extraction of genomic DNA from plant leaf tissue coupled with
amplification and real-time detection. The need for freezing plant tissue in
liquid nitrogen or mechanical disruption and complete genomic DNA
purification has been eliminated. The kits contain all necessary reagents for
both extraction and amplification.
The SYBR Green Extract-N-Amp PCR ReadyMix is the specially formulated PCR
enzyme blend contained in the SYBR Green Extract-N-Amp Plant PCR kit and
SYBR Green Extract-N-Amp Tissue PCR Kit. It is intended to supplement these
kits for researchers who perform more than one amplification per extracted
sample.
Two solutions combine to release genomic DNA from leaf tissue and
neutralize substances inhibitory to PCR. The genomic DNA released into
solution serves as a template for the SYBR Green Extract-N-Amp PCR ReadyMix.
This ReadyMix, which contains SYBR Green fluorescent dye, Taq, JumpStart
antibody, dNTPs and MgCl2, is specially formulated to amplify target
sequences in the cellular lysate.
SYBR® Green Extract-N-Amp™
Plant PCR Kit
Punch leaf tissue sample.
Extract DNA.
15 minutes
Neutralize inhibitors.
Real time PCR.
 sufficient for 100 preparations
XNAPG-1KT
1 kit
S4320-1.2ML
1.2 mL
S4320-12ML
12 mL
S4320-125ML
125 mL
Post-reaction Purification
M
GenElute™ PCR Clean-Up Kit
The GenElute PCR Clean-Up Kit is designed for rapid purification of singlestranded or double-stranded PCR amplification products (100 bp to 10 kb)
from other components in the reaction, such as excess primers, nucleotides,
DNA polymerase, oil and salts. This kit combines the advantages of silica
binding with a convenient spin column format, eliminating the need for
expensive resins or toxic organic compounds such as phenol and chloroform.
1
2
3
1
2
3
1
2
3
2 kb
1.5 kb
750 bp 1 kb
500 bp
300 bp
150 bp
50 bp
M
2 kb
375 bp
143 bp
DNA is bound on a silica membrane within the spin column. The bound DNA
is washed and the clean, concentrated DNA is eluted in the buffer of choice.
Each column can purify up to 100 μL or 10 μg of PCR amplified DNA and
recover up to 95% of PCR products between 100 bp and 10 kb. More than 99%
of the primers and most primer-dimers (<40 bp) are removed.
Purified DNA can be used in enzymatic reactions, conventional or automated
sequencing, cloning and microarray analysis.
• Recovers up to 95% of PCR products between 100 bp and 10 kb
• Purifies up to 100 μL or 10 μg of PCR amplified DNA in 8 minutes
• Removes over 99% of primers and other components
PCR Reaction Components
Comparison of PCR product recovery and primer removal.
Three separate PCR products were purified with the GenElute™ PCR Clean-Up Kit and a comparable kit
from Supplier Q. Products were 143 bp from corn leaf, 375 bp from pBR322, 2 kb from human blood.
Samples were analyzed on a 20% TBE acrylamide gel and visualized by staining with SYBR® Green II
Lanes
1. Unpurified Reaction
2. GenElute™ PCR Clean-Up Kit
3. Supplier Q
 sufficient for 70 purifications
NA1020-1KT
1 kit
GenElute™ 96 Well PCR Clean-Up Kit
1. Prepare Column
Add solution and spin
The purified PCR product is ready for a variety of downstream applications
including sequencing, restriction digest, ligation and microarray analysis.
GenElute 96 Well PCR Clean-Up Kit allows for high throughput purification of
PCR products. The kit provides the necessary reagents for purification of highly
pure PCR products. The DNA recovery is 75-90% for fragments of 100 bp to
10 kb and removes primers, primer-dimers, nucleotides, salts and polymerase.
2. Bind DNA
Spin 1 minute
Once the PCR reaction is complete, the reaction volume is adjusted and
Binding Solution is added. The samples are transferred to the Binding Plate
where the PCR reaction is captured on the silica membrane. The bound PCR
product is washed several times to remove primers, primer-dimers, salts,
nucleotides, and polymerases. Finally the purified PCR product is eluted and
ready for immediate use in downstream applications.
3. Wash Column
Spin 1 minute
Spin 2 minutes
4. Elute DNA
Spin 1 minute
The purified PCR product is ready for a variety of downstream applications
including sequencing, restriction digest, ligation, and microarray analysis.
Pure PCR Product
Sequence was resolved on an ABI 3100 from a purified, 645 bp corn leaf PCR product.
The PCR product was purified with the GenElute™ PCR Clean-Up Kit. The DNA extraction and PCR were
performed using Sigma′s Extract-N-Amp™ Plant PCR Kit. The sequence was obtained by using ABI
BigDye® terminator chemistry and the same primers as the original PCR.
• Complete removal of primers and primer-dimers
• Innovative wash plate minimizes risk of cross-contamination
• Suitable for processing under vacuum and centrifugation
• Time-saving parallel clean-up of PCR products
 sufficient for 1, 96-well plate purifications
PCR9601-1KT
1 kit
 sufficient for 4, 96-well plate purifications
PCR9604-1KT
1 kit
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4
23
9.
kb
6
6.
kb
4
4.
kb
0
2.
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ke
r
GenElute™ Agarose Spin Columns
kb
sigma.com
GenElute Agarose Spin Columns allow for the purification of linear DNA from
agarose gels. The typical recovery is 40 to 45% for 100 bp to 10 kb DNA
fragments. The recovery decreases as the fragment size increases.
The DNA band is excised from an agarose gel and loaded into the spin
column. Embedded within the base of the column are a series of membranes
and filters that hold agarose and impurities back while allowing DNA to
selectively pass through the column into a collection tube during a 10 minute
centrifugation. GenElute Agarose Spin Columns eliminate the need for silicabased resins, DEAE or toxic organic solvents such as phenol and chloroform.
There is no melting, electroelution or enzymatic digestion of the agarose gel.
The purified DNA can be used in most downstream molecular biology
applications including ligation, PCR, restriction digest, cloning, labeling and
hybridization.
• Typical recovery of 40 to 45% for DNA fragments from 100 bp to 10 kb
• No melting or digestion of agarose required
• One-step, 10 minute protocol
Agarose Gel Slice
Recovery of 2 to 23 kb DNA fragments purified using GenElute™ Agarose Spin Columns.
Lambda Hind III markers were pre-heated at 65 °C and underwent electrophoresis on a 2.0% agarose gel.
The 2.0, 4.4, 6.6, 9.4, and 23 kb fragments were excised and purified using GenElute™ Agarose Spin
Columns. Samples were loaded and analyzed on a 2.0% agarose gel.
56500-70EA
70 ea
GenElute™ Minus EtBr Spin Column
GenElute Minus EtBr Spin Columns are based on the GenElute Agarose Spin
Column (56500) and include the incorporation of an additional membrane for
the selective removal of ethidium bromide. Up to 95% of the ethidium
bromide is removed from DNA with a simple 10 minute procedure. The
GenElute Minus EtBr Spin Columns can recover DNA fragments from 100 bp to
10 kb with typical recoveries of 30 to 35%.
1. Load
Place gel slice into
spin column and
spin 10 minutes
2. Collect DNA
The DNA band is excised from an agarose gel and loaded onto the spin
column. The membranes, embedded within the column, retain agarose and
ethidium bromide while allowing DNA to selectively pass through the column
into a centrifuge tube.
The purified DNA can be used in most downstream molecular biology
applications including ligation, PCR, restriction digest, cloning, labeling and
hybridization.
bp
0
bp
60
0
bp
50
bp
0
0
40
bp
30
0
20
0
10
ar
ke
bp
r
Pure DNA
M
38
Recovery of 100-600 bp DNA fragments purified using GenElute™ Agarose Spin Columns.
1 μg of 100 bp DNA ladder underwent electrophoresis on a 2.0% agarose gel. 100, 200, 300, 400, 500,
and 600 bp fragments were excised from the gel, and purified using GenElute™ Agarose Spin Columns
as above. Samples were loaded and analyzed on a 2.0% agarose gel.
• Removes up to 95% of EtBr from DNA
• One-step, 10 minute protocol
• No melting or digestion of agarose required
Electrophoresis was performed on DNA in agarose gels containing ethidium bromide (0.5 μg/mL).
Bands were excised, applied to both the Minus EtBr and Agarose Spin Columns and centrifuged. The
final eluate from each column was compared under a UV light box. The filter within the GenElute™
Minus EtBr Spin Column retained ethidium bromide.
Left Tube: GenElute™ Agarose Spin Column
Right Tube: GenElute™ Minus EtBr Spin Column
56501-70EA
70 ea
 for PCR Purification with Polymerase Removal, Compatible with Rainin
LTS® pipettes
Diffinity RapidTip®
Diffinity RapidTip effectively removes dNTPs, primers and primer dimers while
providing greater than 90% recovery of pure DNA fragments from 100 bp to
10 kb. The functional pipette tip contains everything you need for PCR
purification. The tip is filled with a proprietary adsorption technology that has a
differential affinity for PCR components. The dispensed solution yields purified,
high quality DNA ready for downstream applications such as DNA sequencing,
SNP analysis and microarray printing.
D2947L-8RXN
8 reactions
D2947L-48RXN
48 reactions
D2947L-96RXN
96 reactions
GenElute™ Gel Extraction Kit
The GenElute Gel Extraction Kit combines silica-binding technology with the
convenience of a spin or vacuum column format. DNA fragments of interest
are extracted from slices of an agarose gel and are bound to a silica
membrane. Contaminants are removed by a simple spin or vacuum wash. The
bound DNA is then eluted.
• Single step
• Recovers 90% of high quality dsDNA
• Optimized for 25 μL RXN
The purified DNA is suitable for a variety of downstream applications, such as
automated DNA sequencing, PCR, restriction digestion, cloning, and labeling.
 for PCR Purification
D1947-8RXN
8 reactions
D1947-48RXN
48 reactions
D1947-96RXN
96 reactions
• Bind up to 10 μg of DNA
• Recoveries up to 80%
• Up to 3.5 g can be processed per column
• Compatible with both standard and low-melting agarose in TAE or TBE
buffer
Competitor Q
Sigma
 for PCR Purification, Compatible with Rainin LTS® pipettes
D1947L-8RXN
8 reactions
D1947L-48RXN
48 reactions
D1947L-96RXN
96 reactions
Competitor P
—
2 kb 1.5 kb
————
— 1 kb
— 750 bp
— 500 bp
— 300 bp
— 150 bp
Diffinity RapidTip®2
— 50 bp
Diffinity RapidTip2 effectively removes dNTPs, primers, primer dimers and DNA
polymerase while providing greater than 90% recovery of pure DNA fragments
from 100 bp to 10 kb. The functional pipette tip contains everything you need
for PCR purification with polymerase removal. The tip is filled with a proprietary
adsorption technology that has a differential affinity for PCR components. The
dispensed solution yields purified, high quality DNA ready for downstream
applications such as DNA sequencing, SNP analysis, microarray printing, and
T-A cloning.
• Single step
• Recovers 90% of high quality dsDNA
• Removes polymerase from PCR reactions and cloning
• Optimized for 50 μL reaction
Recovery of 50 bp to 2 kb DNA fragments.
6 μL of the PCR markers (Cat. No. P9577) was electrophoresed on a 1× TBE, 1% agarose gel (Cat.
No. A9414) and each fragment was excised and purified with either competitor Q’s, Sigma’s, or
competitor P’s gel extraction kit per manufacturer’s recommendations. The gel purified samples were
resolved on a 1% agarose gel and the percent recovery was determined via densitometry using a
BIO-RAD Flour-S Imager.
The isolated DNA is suitable for a variety of downstream applications, such as
sequencing, PCR, restriction digestion, cloning and labeling.
NA1111-1KT
 for PCR Purification with Polymerase Removal
D2947-8RXN
8 reactions
D2947-48RXN
48 reactions
D2947-96RXN
96 reactions
1 kit
39
40
sigma.com
SigmaSpin™ Sequencing Reaction Clean-Up
Ideal for removing dye-terminator nucleotides and primers from sequencing
reactions and radiolabeled nucleotides, primers, and fluorescent dyes from
nucleic acid probe labeling reactions.
SigmaSpin™ SigmaSpin™
Post-Reaction
96-Well
Clean-Up
Post-Reaction
Columns
Clean-Up Plates
Sequencing reaction was purified using a SigmaSpin 96-well plate.
pFLAG-MAC™ plasmid (Cat. No. E5644) was sequenced using BigDye® Terminator v3.0 chemistry (20 μL
total reaction volume; BigDye® premix diluted 1:1 with SeqSaver™ [Cat. No. S3938]). Data was generated
on an ABI Prism® 3700 DNA Analyzer with POP-6 polymer and 10× CE Buffer (Cat. No. B4930). PHRED
g20 >600.
 post-reaction clean-up columns
S5059-70EA
1. Prespin
Centrifuge for 2 minutes
@ 750 × g to remove
the buffer
70 ea
 96-well post-reaction clean-up plates
S4309-2EA
2. Load
Load samples into each
of the spin columns
or wells
 10 post-reaction clean-up plates
3. Collect
Centrifuge for 4 minutes
@ 750 × g to collect
purified DNA
 50 post-reaction clean-up plates
S4434-10EA
S4559-25EA
S4559-2X25EA
2 ea
10 ea
25 ea
2 × 25 ea
PCR multiwell plates
Purified DNA
A rigid top plate (included) minimizes plate distortion, assures a dependable fit
with the PCR thermal cycler, and allows for a leak-proof seal with micro-mats
or cap strips. Each well has a maximum capacity of 200 μL.
• Virgin polypropylene
• Fully autoclavable
• Certified DNase- and RNase-free
• Wells have thin walls for rapid temperature equilibration and reduced cycle
time
suitable for (PCR, RT-PCR or DNA purification applications)
 96 well plate for PCR
case = 25
feature ..................................................................................................................... plate format: 96 well standard
Comparison of Sequencing Reaction Clean-Up Methods
Single stranded M13MP18 plasmid was sequenced with a -21M13 forward sequencing primer using ABI
BigDye® Terminator chemistry. Sequencing reactions were resolved on an ABI Prism® 377 XL instrument
with a 48 cm gel cassette containing AutoPAGE™Plus 4.5% acrylamide at 2.88 kV for 7 hrs.
A. Sequencing reactions were precipitated with 70% ethanol and placed on ice for 30 minutes. DNA
pellets were dried and resuspended in TE solution prior to electrophoresis.
B. Sequencing reactions were subjected to post-reaction clean-up with SigmaSpin Post-Reaction CleanUp 96-Well Plates, according to recommended protocol.
SigmaSpin™ Post-reaction Clean-Up technology comes in two convenient forms, 96-well plates and
single spin columns. Each format comes ready for immediate use.
Z374903-2PAK
2 pkg
Related PCR Reagents
Human Genomic DNA
Human Genomic DNA
Human Random Control DNA Panel
Deoxynucleotide Set, 100 mM
Sigma-Aldrich and ECACC have teamed together to provide researchers with
control populations of human genomic DNA for gene regulation and
quantitative PCR research.
The range of Human Random Control (HRC) DNA samples represents a control
population of 480 UK Caucasian blood donors. The HRC DNA is extracted from
lymphoblastoid cell lines derived by Epstein Barr Virus (EBV) that can be
continuously propagated in culture. This ensures an infinite supply of the
unvarying DNA panels. The composition of each panel is completely defined
and standardized so that each lot will be identical. Therefore, the HRC DNA
Panels can be used as Reference Standards as routine quality control in the
laboratory.
The purified HRC DNA is available in 5 different panels (HRC 1 through 5)
consisting of 96 individuals with 2 μg each at a concentration of 100 ng/μL.
The DNA is dissolved in 10 mM Tris (pH 8) 1 mM EDTA.
 Individual dNTPs for routine PCR; 0.25 mL each
0.25 mL each of 100 mM dATP, dCTP, dGTP and dTTP
DNTP100-1KT
 Individual dNTPs for routine PCR; 1 mL each
DNTP100A-1KT
1 kit
Individual Deoxynucleotides
2′-Deoxyadenosine 5′-triphosphate sodium salt solution
NH2
For ordering purposes, the 96-well plate format is listed as a 1 EACH (1EA)
package size.
N
N
O
O
O
O
O
O
NaO P
P
P
OH OH OH
 Panel 1
HRC1-1EA
1 kit
1 ea
N
N
O
OH
 Panel 2
HRC2-1EA
1 ea
 Panel 3
HRC3-1EA
1 ea
 Panel 4
HRC4-1EA
1 ea
 Panel 5
HRC5-1EA
2′-Deoxyadenosine 5′-triphosphate sodium salt solution is used in DNA
sysnthesis reactions such as PCR, DNA sequencing and molecular cloning
techniques.
 100 mM
D4788-25UMO
25 μmol
D4788-.1MMO
0.1 mmol
2′-Deoxycytidine 5′-triphosphate disodium salt solution
1 ea
NH2
N
Deoxynucleotides
O
O
O
HO P O P O P O
ONa ONa OH
Deoxynucleotide Sets
N
O
OH
Deoxynucleotide Set, 10 mM
2′-Deoxycytidine 5′-triphosphate disodium salt is one of the nucleoside
triphosphates used for DNA polymerase driven reactions such as polymerase
chain reaction, DNA sequencing and other molecular biology methods.
 Individual dNTPs for routine PCR; 0.5 mL each
DNTP10-1KT
O
 100 mM (pH 7)
1 kit
D4913-25UMO
25 μmol
D4913-.1MMO
0.1 mmol
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sigma.com
2′-Deoxyguanosine 5′-triphosphate trisodium salt solution
O
10X PCR Buffer
N
HN
• 3Na
H2N
O
O
O
HO P O P O P O
OH OH OH
PCR Buffers
N
 Optimized for routine PCR with MgCl2 included
N
vial = 1.5 mL
O
Composition of the 10× buffer: 100 mM Tris-HCl, pH 8.3 at 25°C; 500 mM KCl;
15 mM MgCl2; 0.01% gelatin
OH
2′-Deoxyguanosine 5′-triphosphate trisodium salt is utilized for methods such
as DNA sequencing, PCR and other DNA polymerase based DNA synthesis and
repair techniques.
 100 mM (pH 7)
D5038-.1MMO
0.1 mmol
P2192-1VL
1 vial
P2192-5VL
5 vials
P2192-5ML
5 mL
P2192-25ML
25 mL
10X PCR Buffer without MgCl2
 Optimized for routine PCR without MgCl2
Thymidine 5′-triphosphate sodium salt solution
10× PCR Buffer for use with Sigma′s PCR enzymes.
Thymidine 5′-triphosphate sodium salt solution is used in DNA synthesis
reactions such as PCR, DNA sequencing, cDNA synthesis and molecular
cloning techniques.
P2317-1VL
P2317-1.5ML
P2317-5ML
1 vial
1.5 mL
5 mL
 100 mM in H2O
T9656-25UMO
25 μmol
T9656-.1MMO
0.1 mmol
 10 mM (pH 7.0)
T7791-.5ML
0.5 mL
10× REDTaq® PCR Reaction Buffer
 To be used with REDTaq® DNA Polymerase
10x REDTaq® PCR Reaction Buffer is a polymerase chain reaction buffer for use
with REDTaq® DNA Polymerase.
B5926-1.5ML
B5926-5ML
1.5 mL
5 mL
Deoxynucleotide Mixes
AccuTaq™ LA 10× Buffer
Deoxynucleotide Mix, 10 mM
The Deoxynucleotide mix is a convenient premixed dNTP solution containing
10 mM each of UltraPure dATP, dCTP, dGTP and TTP sodium salts in high
quality molecular biology grade water. One μL is sufficient for a standard 50 μL
PCR reaction.
Suitable for routine and long PCR, manual and automated DNA sequencing,
cDNA synthesis and labeling reactions.
• Purity of each dNTP: Minimum 99%
• Conveniently formulated; 1 μL is used per 50 μL PCR
• Equimolar amounts of each dNTP means less pipetting
• Minimize risk of contamination in PCR
• UltraPure dNTPs can help maximize consistency and yields in critical PCR
reactions
 Molecular Biology Reagent
D7295-.2ML
D7295-20X.2ML
D7295-.5ML
0.2 mL
20 × 0.2 mL
0.5 mL
 10X Buffer for long and accurate PCR
10× Buffer for AccuTaq LA DNA Polymerase, product code D8045 and D4812
vial = 0.5 mL
B0174-1VL
1 vial
B0174-.5ML
0.5 mL
10X MTP™ Taq Buffer
DNA contaminants can be introduced into PCR through a number of reagents.
To minimize the risk of contaminant DNA during PCR, we offer 10x MTP Taq
buffer to be used with MTP Taq DNA Polymerase (Sigma product D7442). Each
lot of MTP Taq buffer undergoes strict quality control testing to ensure the
absence of contaminating DNA. To prevent false positive PCR results, only
DNA-free reagents should be used in PCR reactions with MTP Taq DNA
polymerase.
 10X buffer, free of DNA contaminants; use with MTP Taq DNA Polymerase
M9943-1.5ML
1.5 mL
Extract-N-Amp™ ReadyMixes
Extract-N-Amp™ ReadyMixes
SYBR® Green Extract-N-Amp™ ReadyMix
SYBR® Green Extract-N-Amp™ PCR ReadyMix™
for analysis.
S4320-1.2ML
R4775-1.2ML
The SYBR Green Extract-N-Amp PCR ReadyMix is the specially formulated PCR
enzyme blend contained in the SYBR Green Extract-N-Amp Plant PCR kit and
SYBR Green Extract-N-Amp Tissue PCR Kit. It is intended to supplement these
kits for researchers who perform more than one amplification per extracted
sample.
1.2 mL
R4775-12ML
12 mL
R4775-125ML
125 mL
1.2 mL
S4320-12ML
12 mL
S4320-125ML
125 mL
PCR Optimizing Reagents
Betaine solution
CH3
H3C N
CH3
5M
O
O-
1.2 mL
The addition of betaine at a final concentration of 0.8-1.6M improves the
amplification of DNA by reducing the formation of secondary structure in
GC-rich regions. The addition of betaine has been reported to enhance the
specificity of the polymerase chain reaction by eliminating the base pair
composition dependence of DNA melting.1
E3004-12ML
12 mL
vial = 1.5 mL
E3004-125ML
125 mL
E3004-1.2ML
REDExtract-N-Amp™ PCR ReadyMix™ for Blood
PCR ReadyMix is intended for use with Sigma′s Extract-N-Amp Blood PCR Kits.
All Extract-N-Amp Kit include a volume of PCR ReadyMix sufficient for at least
one PCR reaction per extraction. However, where additional amplifications are
required, supplemental PCR ReadyMix may be needed. The REDExtract-N-Amp
PCR ReadyMix contains a inert tracking dye that allows for convenient direct
loading of PCR reactions onto agarose gels for analysis.
P8240-12ML
12 mL
Extract-N-Amp™ PCR ReadyMix™ for Blood
PCR ReadyMix™ is intended for use with Sigma′s Extract-N-Amp™ Blood PCR
Kits. All Extract-N-Amp Kits include a volume of PCR ReadyMix sufficient for at
least one PCR reaction per extraction. However, where additional amplifications are required, supplemental PCR ReadyMix may be needed.
P8115-12ML
12 mL
Lit. cited: 1. Rees, W.A., et al., Betaine can eliminate the base pair composition
dependence of DNA melting. Biochemistry 32, 137-144 (1993)
B0300-1VL
1 vial
B0300-5VL
5 vials
Dimethyl sulfoxide
H3C
O
S
CH3
Supercools easily and remelts slowly at room temperature. Solidified product
can be re-liquified by warming to room temperature without detriment to the
product.
Dimethyl sulfoxide (1-10%) has been shown to accelerate strand renaturation
and is believed to give the nucleic acid thermal stability against depurination.
As a PCR cosolvent, DMSO may help improve yields, especially in long PCR.
vial = 1 mL
D9170-1VL
1 vial
D9170-5VL
5 vials
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Magnesium chloride solution
Water
 25 mM MgCI2 solution for PCR
H2O
Suitable for optimization of polymerase chain reactions.
Suitable for polymerase chain reaction (PCR)
vial = 1.5 mL
vial = 1.5 mL
concentration ............................................................................................................................................ 25 mM±1 mM
M8787-1VL
1 vial
M8787-5VL
5 vials
M8787-5ML
5 mL
W1754-1VL
1 vial
W1754-5VL
5 vials
Post-reaction Analysis Reagents
Single-strand Binding Protein from Escherichia coli
DNA Ladder Markers
 Single strand binding protein to enhance PCR specificity
Binds with high specificity to single-stranded DNA. Can be useful in enhancing
the specificity of PCR and in enabling the sequencing of problematic DNA
templates.
Solution in 20 mM Tris-HCl, pH 8.0, 0.5 M NaCl, 0.1 mM EDTA, 0.1 mM DTT,
50% glycerol
S3917-.1MG
0.1 mg
S3917-.5MG
0.5 mg
2,000
600
900
700
600
500
Glycerol
400
300
200
100
50
OH
HO
1,000
900
800
700
3,000
OH
1,000
800
500
400
450
350
250
150
300
200
Glycerol is used both in sample preparation and gel formation for
polyacrylamide gel electrophoresis. Glycerol (5-10%) increases the density of a
sample so that the sample will layer at the bottom of a gel’s sample well.
Glycerol is also used to aid in casting gradient gels and as a protein stabilizer
and storage buffer component.
G5516-100ML
100 mL
G5516-500ML
500 mL
G5516-1L
1L
10,000
8,000
6,000
5,000
4,000
3,000
2,500
2,000
1,500
1,000
500
100
D3812 on a 1.5% agarose gel
D3687 on a 2% agarose gel
DirectLoad™ Step Ladder, 50 bp
Contains 17 fragments 50-500 bp by 50, 600-900 bp by 100, and 1000-3000 bp
by 1000.
Ribonuclease inhibitor human
This inhibitor of RNase activity is isolated from human placenta and supplied as
a buffered aqueous glycerol solution.
Useful for in vitro inhibition of ribonucleases, including procedures like cDNA
synthesis, RT-PCR, and in vitro transcription and translation.
Suitable for fragment size determination using DNA electrophoresis. The
DirectLoad™ marker comes ready-to-use in a gel loading solution. Can be used
with either agarose or polyacrylamide gels; the recommended agarose gel
concentration is 2.0%.
Solution in 50% glycerol, 20 mM HEPES-KOH, pH 7.6, 50 mM KCl and 8 mM DTT
Solution in 5% glycerol, 4.2 mM EDTA, 0.083% orange G and 0.0125% xylene
cyanol
 Inhibits ribonucleases during transcription and translation experiments
sufficient for 100 loads
concentration ........................................................................................................................ 30,000-50,000 units/mL
R2520-2.5KU
2500 units
R2520-10KU
10000 units
R2520-20KU
20000 units
D3812-1VL
1 vial
Post-reaction Analysis Reagents: DNA Ladder Markers
DirectLoad™ PCR 100 bp Low Ladder
DirectLoad™ Wide Range DNA Marker
Contains 10 fragments, 100–1,000 bp in exact 100 bp repeats.
Suitable for fragment size determination using DNA electrophoresis on
agarose or polyacrylamide gels. The DirectLoad™ marker comes ready-to-use
in a gel loading solution with tracking dye. The recommended agarose gel
concentration is 2.0%
Solution in 5% glycerol, 4.2 mM EDTA, 0.09% orange G, 0.0125% xylene cyanol
sufficient for 75 loads
concentration ................................................ 4.0% agarose in TAE Buffer (for DNA electrophoresis)
D3687-1VL
1 vial
DirectLoad™ 1 kb DNA Ladder
Contains 16 fragments, 50–10,000 bp which produce logically spaced bands
on an agarose gel.
DirectLoad™ marker comes ready-to-use in a gel loading solution. This marker
can be used with either agarose or polyacrylamide gels. The recommended
agarose gel concentration is 0.75% for the larger fragments and 2.0% for the
smaller fragments.
DirectLoad marker comes ready-to-use in a gel loading solution for use with
either agarose or polyacrylamide gels. The recommended agarose gel
concentration is 0.75% for the larger fragments and 2.0% for the smaller
fragments.
Solution in 2.5% Ficoll (Type 400), 0.0125% bromophenol blue, 0.00625%
xylene cyanol
 1 vial sufficient for 100 loads
Contains 11 fragments consisting of 500 bp repeats from 0.5 to 3 kb, 1 kb
repeats from 3 to 6 kb and 2 kb repeats from 6 to 10 kb.
DirectLoad™ marker comes ready-to-use in a gel loading solution. This marker
can be used with either agarose or polyacrylamide gels. The recommended
agarose gel concentration is 0.75% for this marker.
DirectLoad marker comes ready-to-use in a gel loading solution for use with
either agarose or polyacrylamide gels. The recommended agarose gel
concentration is 0.75%.
sufficient for 100 uses
D7058-1VL
1 vial
1 kb Ladder
Contains 11 fragments consisting of 500 bp repeats from 0.5 to 3 kb, 1 kb
repeats from 3 to 6 kb and 2 kb repeats from 6 to 10 kb.
Solution in 2.5% Ficoll® (Type 400), 0.0125% bromophenol blue, 0.00625%
xylene cyanol
Suitable for size determination of double-stranded DNA using DNA electrophoresis. This marker can be used with either agarose or polyacrylamide gels.
The recommended agarose gel concentration is 0.75% for this marker. A
sample of the marker should be diluted with gel loading buffer to an
appropriate loading concentration.
D3937-1VL
Solution in 10 mM Tris-HCl, pH 8.0, and 1 mM EDTA
1 vial
10,000
8,000
6,000
4,000
3,000
2,000
1,550
1,400
1,000
750
500
D0428-1VL
10,000
8,000
6,000
5,000
4,000
3,000
2,500
2,000
1,500
1,000
400
300
DNA Ladder, Supercoiled
Contains 11 supercoiled fragments, 2,067–16,210 bp
16,210
14,174
12,138
10,102
8,066
7,045
6,030
5,012
3,990
2,972
200
100
50
2,067
500
1 vial
Especially designed for size determination of naturally supercoiled DNA using
DNA electrophoresis. The recommended agarose gel concentration is 0.7% for
this marker. A sample of the marker should be diluted with gel loading buffer
(G2526) to an appropriate loading concentration. Typically 0.2 μg per well
(0.02 μg/μl, 10 μl load) is sufficient to be seen using ethidium bromide staining.
Especially designed for size determination of naturally supercoiled DNA using
electrophoresis. The recommended agarose gel concentration is 0.7% for this
marker.
Solution in 10 mM Tris-HCl, pH 8.0, 1 mM EDTA
An aliquot of the marker should be diluted with gel loading buffer (G2526) to
an appropriate loading concentration before use. Typically 0.02 μg/μL with a
10 μL load is sufficient to be seen using ethidium bromide staining.
D5292-10UG
10 μg
D5292-25UG
25 μg
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1,000
900
800
700
1,000
900
800
700
600
500
600
500
400
1,000
900
800
700
400
300
600
300
Especially designed for size determination of PCR generated DNA fragments.
Mixing the two markers together will produce a band every 20 bp, with every
fifth band more intense. These markers can be used with either agarose or
polyacrylamide gels. The recommended agarose gel concentration is 2.5% for
these markers. A sample of each marker should be diluted with the supplied
gel loading buffer to the desired loading concentration.
200
500
200
PCR Low Ladder Set
The set provides separate tubes of the 20 bp and 100 bp ladders (25 μg each).
Includes 6× loading buffer and an empty tube for storing the mixture.
400
D7808-1SET
100
300
1 set
100
200
3,000
2,000
20
100
20
4,182
P1598 on a 2.5% agarose gel
P1473 on a 2% agarose gel
1,000
2,000
PCR 20 bp Low Ladder
1,000
900
800
700
750
600
500
500
450
350
300
1,500
2,337
2,091
1,845
1,599
1,353
1,230
1,107
984
861
739
615
Contains 50 bands, 20–1,000 bp in exact 20 bp repeats.
Especially designed for size determination of PCR-generated DNA fragments.
This marker can be used with either agarose or polyacrylamide gels. The
recommended agarose gel concentration is 2.5% for this marker. A sample of
the marker should be diluted with gel loading buffer to the desired loading
concentration.
300
50
369
250
200
150
492
150
246
100
123
P1598-40UG
50
40 μg
P9577 on a 2% agarose gel
S7025 on a 2% agarose gel
PCR 100 bp Low Ladder
Contains 10 bands, 100–1,000 bp in exact 100 bp spaced (ladder) recombinant
repeats.
recommended agarose gel concentration is 2.0% for this marker. A sample of
the marker should be diluted with gel loading buffer to the desired loading
concentration.
123 bp Ladder
Contains 35 fragments, 123 - 4182 bp
Each fragment contains between 1-34 repeats of a 123 bp region of the rat
prolactin gene. This marker can be used with either agarose or polyacrylamide
gels. The recommended agarose gel concentration is 1.5% for this marker. A
sample of the marker should be diluted with gel loading buffer to an
appropriate loading concentration. Up to 25 bands may be resolved at once
on agarose or polyacrylamide gels.
Solution in 10 mM Tris-HCl, pH 7.5, 0.1 mM EDTA, 50 mM NaCl
P1473-1VL
1 vial
Dilute the marker with gel loading buffer to an appropriate loading
concentration.
D5042-.1MG
0.1 mg
D5042-250UG
250 μg
Post-reaction Analysis Reagents: DNA Ladder Markers
PCR Marker
RNA Markers
Contains 8 bands, 50–2,000 bp in predictably spaced (ladder) double-stranded
DNA fragments.
10,000
6,000
recommended agarose gel concentration is 2.0% for this marker.
4,000
3,000
2,000
A vial of 6× gel loading buffer is included for diluting samples.
Vial contains 250 μl
1,500
6,583
4,981
3,638
1,000
2,604
1,908
1,383
Components
PCR Marker 250 μL
P9577-1VL
955
623
500
1 vial
281
200
Step Ladder, 50 bp
R7020 on a 1% agarose gel with formaldehyde
R7644 on a 1% agarose gel with formaldehyde
Contains blunt-ended fragments of DNA, 50-500 bp by 50, 600-900 bp by 100,
and 1000-3000 bp by 1000 (17 bands).
This marker is especially designed for size determination of PCR generated
DNA fragments and can be used with either agarose or polyacrylamide gels.
The recommended agarose gel concentration is 2.0% for this marker. A sample
of the marker should be diluted with gel loading buffer to an appropriate
loading concentration.
Solution in 10 mM Tris-HCl, 10 mM NaCl, 1 mM EDTA, pH 7.5
S7025-50UG
50 μg
Transcript RNA Markers 0.2-10 kb
Contains nine defined RNA transcripts, 200–10,000 nucleotides
Suitable for size determination of small RNA using native or denaturing
agarose gel electrophoresis. The recommended agarose gel concentration is
1.0%. A sample of the marker should be diluted with gel loading buffer (R4268)
to an appropriate loading concentration. Typically, 2 μg per well is sufficient to
be seen with ethidium bromide staining.
R7020-50UG
50 μg
Transcript RNA Markers 0.28-6.6 kb
Contains a mixture of nine RNA transcripts, 281–6,583 bases
Suitable for size determination of small RNA using native or denaturing
agarose gel electrophoresis. The recommended agarose gel concentration is
1.0%. A sample of the marker should be diluted with gel loading buffer
(R 4268) to an appropriate loading concentration. Typically, 3 μl per well is
sufficient to be seen with ethidium bromide staining.
Solution in 10 mM Tris-HCl, pH 8.0, with 1 mM EDTA
R7644-1VL
1 vial
RNA Marker template set
 ~0.5 mg/mL in 10 mM Tris-HCl, pH 7.5, 1 mM EDTA
A mixture of seven linearized plasmids for use as templates for RNA markers.
In vitro transcription with T7 RNA polymerase produces transcripts 100, 200,
300, 400, 600, 800 and 1,000 bases long. The transcripts can be labeled by
including biotinylated, radioactive, or other labeled rNTP′s in the reaction. The
markers are useful for RNase protection assays or for electrophoresis of low
molecular weight RNA fragments.
R4142-5UG
5 μg
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sigma.com
PCR Accessories
Thermal Cyclers
Plates
Labnet MultiGene™ OptiMax thermal cycler
The new MultiGene OptiMax brings new speed and features to our Thermal
Cycler line. It utilizes an easy user interface, having a much greater speed of
operation by increasing the ramping speed and eliminates overshooting or
undershooting with contributes to longer run times. New features like PC
Viewer and a TM calculator allows you to connect to a Windows based PC
using a USB interface and view the actual temperature profiles in real time.
The new 6-block gradient format allows you to select the temperatures you
wish to optimize rather than have the system select for you.
• No condensation after overnight cooling at 4 °C
• 6-segment block temperature control with users able to select 6-segment
temps independently
• TM Calculator
• PC Viewer
• Simple user interface
200 complete programs in memory, 6 temperature blocks in 4 x 4 well format,
Peltier heating/cooling method
PCR multiwell plates
A rigid top plate (included) minimizes plate distortion, assures a dependable fit
with the PCR thermal cycler, and allows for a leak-proof seal with micro-mats
or cap strips. Each well has a maximum capacity of 200 μL.
• Virgin polypropylene
• Fully autoclavable
• Certified DNase- and RNase-free
• Wells have thin walls for rapid temperature equilibration and reduced cycle
time
suitable for (PCR, RT-PCR or DNA purification applications)
 96 well plate for PCR
case = 25
feature ..................................................................................................................... plate format: 96 well standard
Z374903-2PAK
2 pkg
 384 well PCR plate, polypropylene, skirted, non-sterile, 50/cs
384 well PCR plates are skirted for compatibility with automation systems.
Wells have raised rims to ensure contact with sealing film and reduce
evaporation. Each well has a capacity of 40 μL and a working volume of 25 μL.
Multigene OptiMax
case = 50
feature ................................................................................................................... plate format: 384 well standard
Z374911-1PAK
Multigene OptiMax with lid open
Cat. No.
Z759155-1EA
Z759163-1EA
Temp. Range (°C)
4-99.9
AC
120 V
4-99.9
240 V (European 2-pin plug)
1 pkg
PCR Accessories
Plates
 96 well plate, Thermowell GOLD PCR plate, half skirt, polypropylene,
conical bottom, clear, non-sterile, 50/cs
Greiner 96 well plates, polypropylene
PCR Plates have a standard microplate footprint and are compatible with
thermocyclers of most major brands. DNAse and RNAse free and nonpyrogenic by LAL method to <0.06 EU/mL.
feature ..................................................................................................................... plate format: 96 well, half skirt
CLS3753-50EA
Use for polymerase chain reaction; Free of detectable DNase/RNase, human
DNA and pyrogens.
50 ea
Corning® Thermowell GOLD PCR 384 well plates
• 384 well PCR microplates feature a rigid construction that is ideal for highthroughput (HT) automation
• Suitable for PCR, DNA sequencing and RT-PCR
• Uniform wall thinness assures efficient heat transfer while minimizing wellto-well variability
• 100% tested for well integrity and certified DNase- and RNase-free
• Nonsterile, without lids
case = 40
suitable for (high-throughput (HT) applications; PCR, DNA sequencing and
RT-PCR)
case = 50
 96 well, half skirted
96 well microplate, without lid, half-skirt design; alphanumeric well coding,
length: 126.24 mm; width: 86 mm.
96W PCR Microplate, well working volume 200 μL; total well volume 250 μL.
Z711055-40EA
pack = 10
feature ................................................................................................................... plate format: 384 well standard
lid .................................................................................................................................................................................................. No
40 ea
 96 well, full skirted
96 well microplate, without lid, fμLl-skirt design; alphanumeric well coding,
length: 127.76 mm; width: 85.34 mm.
96W PCR Microplate, well working volume 150 μL; total well volume 200 μL.
Z711063-40EA
40 ea
• 96 well PCR microplates with elevated skirts feature a rigid construction that
is ideal for automation
• Clear medical grade polypropylene is suitable for RT-PCR
• Suitable for PCR, DNA sequencing and RT-PCR
• Uniform wall thinness assures efficient heat transfer while minimizing wellto-well variability
• 100% tested for well integrity and certified DNase- and RNase-free
• Nonsterile, without lids
suitable for (PCR, DNA sequencing and RT-PCR)
case = 50
pack = 10
 384 well plate, Thermowell GOLD PCR plate, polypropylene, conical
bottom, clear, non-sterile, 50/cs
This is the suggested replacement for CLS3735
CLS3757-50EA
50 ea
Micro mats for PCR plates
•
•
•
•
•
•
•
•
Fits most popular 96-well PCR plates
Minimized evaporation in PCR and storage
Alpha numerics printed on each mat, for easy well identification
Pierceable with syringe
Not compatible with DMSO and other harsh solvents
Tabs for easy application/removal of mat
Reusable for up to 5 times
When used with a screw- or clip-down thermal cycler lid, provides 100%
sealing
• Fully autoclavable, reversible
Z374938-5EA
5 ea
Z374938-10EA
10 ea
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AlumaSeal® CS Films for cold storage
Plate Seals
AlumaSeal® 96 film
 thick aluminum foil sealing film for use with 96 well plates
A 38 μm thick aluminum sealing foil film with a 38 μm layer of acrylic adhesive
for use with 96 well plates. Fits inside the rim of raised-rim plates. AlumaSeal 96
sealing films have one partial-width end tab with no perforations.
•
•
•
•
•
Film dimensions are 127 mm x 78 mm (9.5 mm end tab)
Available as non-sterile only
Pierceable
Recommended temperature range between: -80 °C to +120 °C
Certified DNase-, RNase-, and nucleic-acid-free
AlumaSeal CS sealing films are a specially formulated aluminum foil with a
50 μm layer of acrylic adhesive that withstands cold storage at temperatures
between: -80 °C to +120 °C . Unlike other sealing films in the AlumaSeal group,
AlumaSeal CS sealing films are not recommended for PCR or thermocycling.
A single non-perforated end tab simplifies application. Simply hold the tab and
strip the backing from the body of the sealing film as it lays on the plate to
avoid curling.
•
•
•
•
Film dimensions are 82.6 mm x 132.6 mm (9.5 mm end tab)
Excellent barrier properties to retard evaporation
Easily pierceable with pipet tips or robotic probes for sample recovery
Certified DNase-, RNase-, and nucleic-acid-free
Z722634, Z722642
 formulated aluminum foil sealing films
Z722634-100EA
Z721549-100EA
100 ea
AlumaSeal® II film
 for PCR and cold storage
A 38 μm soft non-permeable aluminum foil sealing film with strong medicalgrade adhesive, AlumaSeal® II sealing films eliminate the need for heat-sealing
devices or mats during thermal cycling. Each sealing film measures 82.6 x
142.9 mm and offers sufficient sealing area for all PCR plates. Length between
the perforations with end tabs removed is 125.4 mm. Compared to other
aluminum foils, AlumaSeal® II has less tendency to roll back or itself when
removing the backing paper and conforms well to the plate during
application. Sterile product is packaged in 100 μm tamper-evident bags of 25.
•
•
•
•
Easily pierceable with single or multichannel pipettors and robotic probes
Heat & cold resistant, recommended for temperatures from -80 °C to +120 °C
Certified DNase, RNase, and Nucleic Acid free
Excellent barrier properties, virtually no sample evaporation or drying
pack = 50 ea (pkg of 50ea in two zip bags of 25ea)
Z721530-50EA
50 ea
100 ea
SealPlate® film
Sheets pre-cut to fit standard multiwell plates. Both film and adhesive are inert
and compatible with microplate procedures. Adhesive forms a tight,
waterproof seal, preventing cross-contamination and evaporation. SealPlate
film is 2 mil (0.05 mm) polyester, with a functional temperature range of -40 to
95 °C.
Cat. No.
Z369659-100EA
Z369667-100EA
Sterile
No
Yes
PCR Accessories
Plate Seals
Greiner multiwell plate sealers
ThermalSeal A™ sealing films
Greiner microplate sealers are RNase free, DNase free and nonpyrogenic. They
feature a temperature tolerance range of -70 °C up to 110 °C.
 SILVERseal™ sealing film, adhesive aluminium foil on carrier paper,
piercable, silver
Note: Pierceable aluminum foil coated with an acrylate adhesive.
•
•
•
•
Temperature resistant from -80 °C to 110 °C
Ideal for PCR applications
Ideal for storage of sample material and active agents
L: 80 mm x W: 140 mm
Material:
Foil: Aluminum
Glue: Acrylate Adhesive
Release Liner: Polyethylene-coated paper (with silicone coating on one side)
suitable for (for coverage/ storage of microplates; PCR applications)
 with improved adhesion
ThermalSeal A sealing films are an advanced version of Excel Scientific′s classic
ThermalSeal A sealing films for PCR. ThermalSeal A films consist of 50 μm thick
clear polypropylene with a thicker 50 μm layer of a stronger adhesive
formulated for improved protection against evaporation and leakage during
thermal cycling. Intended for non-real-time PCR applications, ThermalSeal A
films are designed to fit inside the edge of raised-rim PCR plates and provide
more secure sealing of all wells because the center of the film does not extend
over the plate rim. Two end tabs allow for easy positioning of the film on the
plate but are perforated for easy removal if necessary.
Each film measures 77.8 x 135.5 mm overall. Length between the perforations
with end tabs removed is 118.1 mm. Each package of ThermalSeal A films
includes one sealing paddle for ensuring a firm seal, also compatible with
raised-rim plates.
•
•
•
•
•
•
Recommended for non-real-time PCR thermocycling applications
Stronger and thicker 50 μm adhesive layer for secure seal
Heat resistant, recommended for temperatures from -40 °C to +125 °C
Can be used with raised-rim PCR plates and standard plates
Two end tabs, perforated for easy removal
Each package of ThermalSeal A™ films includes one sealing paddle to ensure
a firm seal
• Certified DNase, RNase, and Nucleic Acid free
pack = 100
pack = 100
Z617601-100EA
100 ea
ThermalSeal® films, classic
A 50 μm heat-resistant polypropylene sealing film with a 25 μm layer of acrylic
adhesive designed for thermal cycling applications. Each sealing film measures
79.4 x 135.1 mm and offers sufficient sealing area for all PCR plates. Length
between the perforations with end tabs removed is 123.1 mm. ThermalSeal
sealing films are not pierceable. For applications where piercing with pipet tips
or robotic probes is required for product recovery, use AlumaSeal® sealing
films. For real-time PCR applications where maximum optical clarity is required,
use ThermalSeal RT sealing films.
• heat resistant, recommended for temperatures from -40 °C to +120 °C
• easier to apply than aluminum foils, no tendence to roll back
• certified DNase-, RNase-, and nucleic-acid-free
Z369675, Z369683
Cat. No.
Z369675-100EA
Z369683-100EA
Sterile
No
Yes
Z723304
Z723304-100EA
100 ea
51
52
sigma.com
ThermalSeal RTS™ Sealing Films
ThermalSeal® Films for Real-time PCR
ThermalSeal RT™ film
A combination of optically transparent polyester film with a strong, ultrasmooth, non-absorbing, non-fluorescing medical-grade adhesive for superior
performance in real-time PCR applications. In place of the customary paper
backing, ThermalSeal RT has a plastic liner which is easily removed before use
and contributes smoothness and extreme optical clarity to the adhesive.
Dimensions are 3.125 x 5.625 in. to seal PCR plates. Length between the
perforated ends is 4.8 in.
• Ultra-high optical clarity
• Heat resistant, recommended for temperatures from -40 °C to +120 °C
• Certified DNase, RNase, and nucleic acid free
•
•
•
•
•
•
•
•
•
High optical clarity
Minimal to no autofluorescence
Chemically inert; no extractables except at extreme pH
DMSO resistant for HTS
Heat resistant, recommended for temperatures from
-70 °C to +100 °C
Fit within raised plate rim to prevent loss of seal due to film lifting
Silicone adhesive forms the strongest available seal for evaporation
prevention
• Non-tacky adhesive layer simplifies handling of film prior to sealing
Z734438
ThermalSeal RT film
Z707465-100EA
 for qPCR, storage & crystallization
100 ea
Z734438-100EA
ThermalSeal RT2RR™ film
 sealing films for real-time qPCR
ThermalSeal RT2RR™ 50 μm polyester sealing films for real-time PCR are sized
to fit within the edges of raised-rim 96 well plates. The same consistent ultrahigh optical clarity as Excel′s ThermalSeal RT™ sealing films makes possible
more reproducible, reliable, and consistent DNA amplification measurements.
An inert, strong, temperature-resistant adhesive assures reliable sealing around
each well. Two end tabs assist in positioning the sealing film. Easy removal of
the end tabs at perforated boundaries prevents lifting and higher evaporation
rates that can occur with sealing films that overlap the plate rim.
Recommended for specific DNA sequence detection in clinical diagnostics,
forensic science and basic research.
Dimensions 77.8 by 130.8 mm. With end tabs removed, length is 118.1 mm
with 45° corners.
•
•
•
•
•
pack = 100 ea
Ultra-high optical clarity
Fit within plate rim to prevent evaporation due to sealing film lifting
Heat resistant, recommended for temperatures from
-40 °C to +120 °C
PurePak PCR Microtubes
PurePak PCR microtubes
Reaching into a bulk bag of tubes can cause contamination; PurePak
packaging solves this problem by dividing tubes into ten separate PurePaks.
PurePaks can be opened as needed to protect unused tubes from
contamination. Thin walled tubes are precision-molded with premium, nonwettable polypropylene and receive multi-point, quality inspections to ensure
unsurpassed performance. Certified Rnase-, DNase- and pyrogen-free.
thin walls
pkg = 1,000 tubes
 0.2mL PCR tubes, nuclease and pyrogen-free
case = 10 packs
P3114-1PAK
1 pkg
P3114-1CS
1 case
 0.5 mL PCR tubes, nuclease and pyrogen-free
pack = 100 ea
P3364-1PAK
Z722553-100EA
100 ea
100 ea
1 pkg
PCR Accessories
PCR Microtubes
PCR microtube and cap strips
PCR Microtubes
PCR microtubes with attached caps
Thin-walled for efficient thermal transfer and shorter cycle times; fits all leading
thermal cyclers including PerkinElmer, Biometra, MJ Research, Techne, Grant
and Strategene (0.65 mL only). Certified DNase- and RNase-free. All tolerate
organic solvent reactions and temperatures from -4 to 121 °C.
Strips of eight tubes connected with double bridges to avoid accidental
separation. Caps also are in strips of eight. Can be cut apart to use individually
if desired.
pkg = 250 strips (8 tubes or caps per strip)
Z374962-1PAK
Autoclavable
case = 4 pkg
pkg = 250 tubes
Z374873-1PAK
1 pkg
Z374873-1CS
1 case
Z374881-1PAK
1 pkg
Z374881-1CS
1 case
Scale-up Your Experiment.
Milligram to gram quantity custom oligos from
Sigma® Life Science, for when you need more.
Bioflexible.
Choose Sigma for impeccable quality and rapid delivery of milligram to
gram quantities of DNA and RNA oligos. Sigma’s iScale Oligos™ are perfectly
suited for in vivo experiments, high throughput assays, structural studies or
commercial manufacturing — all from the name you trust.
Order milligram quantities, sigma.com/iscaleoligos
1 pkg
53
54
sigma.com
The Sigma Advantage
Global Manufacturing
Sigma is the world’s leading supplier of custom oligonucleotides and peptide
libraries for the global life science research community. Along with cuttingedge technology, competitive prices and continuous investment in worldwide
operations, we believe the following sets us apart:
Sigma has manufacturing sites in 9 countries: Australia, Canada, Germany,
India, Israel, Japan, Singapore, UK, and USA. Our global customers receive
consistent high-quality products, no matter where their research happens.
•
•
•
•
•
OEM and Contract Manufacturing
Expertise of our Scientists
Outstanding Customer Service & Technical Support
Commitment to Quality
Global Manufacturing
OEM and Contract Manufacturing
Expertise of Our Scientists
With over 25 years experience in oligonucleotide synthesis, we have the
expertise to meet your project requirements. Our scientists collaborate with
researchers from around the world in developing new products and
technologies. We meet customers’ specifications, no matter how complex.
Outstanding Customer Service & Technical
Support
Our dedicated staff of highly trained service specialists is available by phone
and email to provide timely solutions to every inquiry. Sigma customers are
provided with best-in-class service and support including:
•
•
•
•
Primer & Probe Design Services
Applications Support
Troubleshooting Assistance
Real-time Order Status
Our Commitment to Quality
We analyze both in-process and final product quality to meet and exceed the
high standards of researchers. Sigma’s state-of-the-art analytical instruments,
coupled with a robust quality management system, enable Sigma to
guarantee product performance through:
• Effective Quality Management Systems
• Document and Record Controls
• Effective External and Internal Audits, Corrective and Preventive Actions, and
Training Programs
• Vendor Management
Delivery, quality, expertise and reliability make Sigma a long-term, trusted
partner for all oligonucleotide and PCR reagent needs.
• Project Management
• Product & Process Development Support
• Custom Formulations & Packaging
Custom Oligonucleotides: DNA
Oligos in Plates for High-throughput Applications
Custom Oligonucleotides
DNA
Oligonucleotides are at the foundation of modern life science research.
Whatever your application, Sigma provides the breadth of product and
superior quality and service, enabling your next great discovery.
Options
0.025, 0.05, 0.2, 1.0, 10 and 15 μmole
Desalt, Cartidge, HPLC, PAGE
10–110 bases
Phosphodiester/Phosphorothioate
Amine, Thiol, Phsophorylation, PEG Spacers, 2′OMe-RNA, Biotin, etc.
Fluorescein, 6-FAM™, TAMRA™, JOE™, Rhodamine™ dyes, Texas Red®, WellRED (D2,
D3 & D4), HEX™, TET™, Cy® dyes
BHQ1™, BHQ2™, Dabcyl
MALDI-TOF MS, Electrospray Ionization MS, OD by UV Spectroscopy
Duplicate Labels, Technical Datasheet
Dry or In-Solution (normalized)
2 mL–50 mL Tubes, Microtiter Plates
Non-fluorescent Dyes:
Quality Control:
Labels & Documentation:
Format:
Packaging:
Primers Made Easy
We offer a range of DNA oligonucleotides when the focus is speed,
convenience and simplicity. These primers include technical datasheet
documentation, extra product labels, and quick delivery.
DNA Oligo Options
Product
E@sy Oligo™
Pure & Simple
Purification
Desalt
Cartridge
1. Approximately 15 nmoles or 3 OD
Options include:
Oligos in Plates for High-throughput Applications
DNA Oligo Specifications
Criteria
Scale:
Purification:
Length:
Backbone:
Common Modifications:
Fluorescent Dyes:
Whether your application requires a few or more than 100,000 oligos, Sigma
provides the format to best suit your application, automation and LIMS system.
We conduct shipping validation studies on all plate and seal combinations. Let
our experts recommend the right plate for your system.
Length
15–30 mers
12–35 mers
Format
(~15 nmoles)
100 μM in water
Dry1
Shipment
Next Day
Next Day
Criteria
Plates:
Options
96 or 384
Deep well or shallow well
Matrix plates
Micronic plates
Daughter plates (copies) for all formats
Peelable and pierceable heat seals
Plate Seals:
Concentrations and Volumes: Variety of concentration options available with fixed or variable volumes
Single oligo per well
Formulations:
Forward & reverse primers in a single well
Complex formulations containing 2–20 oligos
Plate position verification by mass spectrometry
Quality Control:
Deionized water
Buffers:
TE Buffer
Customer-specified buffers
1-D or 2-D barcodes
Labels & Documentation:
Customized Quality Assurance Datasheet
Sigma employs liquid-handling systems to normalize oligonucleotide
concentrations within a narrow range. Our investment in automation enables
quick delivery of customer-specified products, no matter how complex.
Contact your local Sigma office to discuss your project and request a
quotation.
iScale Oligos™ DNA
iScale Oligos are perfect for in vivo, high-throughput and diagnostic projects
for which larger amounts are required.
iScale Oligos™ Options
Criteria
Amounts Offered:
Purification:
Quality Control:
In vivo Options:
Modifications:
Options
10, 25, 50, 100, 250, 500 mg (Inquire for larger quantities)
Desalt, RP-HPLC, IE-HPLC
Mass Spectrometry, including LC-MS, HPLC
Dialysis, filtration, and endotoxin testing
Extensive offering of dyes & quenchers, amines, biotins, linkers, etc.
Sigma can provide a custom solution for every project guaranteed!
55
56
sigma.com
qPCR Probes and Assay Design Services
Assay Design
Quantitative Real-time PCR Probes
Sigma offers the largest licensed probe selection for your qPCR needs.
Our offering includes:
•
•
•
•
•
Sigma is pleased to offer OligoArchitect for all of your primer and probe design
requirements. OligoArchitect includes both our complimentary online design
tool and our unique consultative service.
Dual-labeled Probes
Molecular Beacons
LightCycler® Probes
Scorpions® Probes
Locked Nucleic Acids (LNA)-containing Probes
OligoArchitect Online
Sigma’s probes are provided in a format to simplify your experimental
planning.
Product features include:
•
•
•
•
•
•
•
Amounts: 1, 3, 5, and 10 OD
Purification: HPLC
Sequence Lengths: 15 - 40 bases
Modifications: LNA available
Dyes: Comprehensive offering of reporters and quenchers
Quality: 100% mass spectrometry
Format: Supplied dry in amber tubes
Theoretical qPCR Probe Yields
Guaranteed
OD Yield
1
3
5
10
Approximate
No. of nmoles*
4
12
20
40
Approximate
No. of μg
32
96
160
320
OligoArchitect Primer and Probe Design Services
Approximate
No. of Reactions*
800
2,400
4,000
8,000
For routine needs, improve your assay with our OligoArchitect online design
tool powered by the industry standard Beacon Designer (Premier Biosoft). The
user-friendly interface utilizes the latest algorithms, provides results in real time,
supports templates up to 10,000 base pairs, and allows for the adjustment of
input parameters such as homopolymer run/repeat maximum length, G/C
clamp length, and maximum primer pair Tm mismatch. All reported
sequences, associated properties, and assay parameters are available for export
to Excel and convenient ordering.
OligoArchitect Consultative
For more complex and demanding applications, utilize our consultative service
to ensure the success of your assay. With personal consultation from our
expert, technical support scientists, your request, including all sequences and
data analysis, will be MIQE compliant and returned to you within 24 hours. If
required, you will also receive follow-up assay optimization, data analysis
assistance, and troubleshooting support.
Powered by Beacon Designer™ from PREMIER Biosoft, OligoArchitect™ Online
is the most comprehensive qPCR assay design tool available.
Try our online design tool at sigma.com/probedesignonline
*Estimate is based on 4 nmoles or 32 μg for 1 OD and 200 nM in 25 μL reaction (5.0 pmol/reaction).
Estimate is based on average sequence length of 25 bases.
Trademarks
The following trademarks and registered trademarks are accurate to the best of our knowledge at the time of printing. Please consult
individual manufacturers and other sources for specific information.
Agilent Technologies, Inc. — Mx3000P®, Mx3005P®, Mx4000™
Applera Corporation or its subsidiaries in the US and/or certain other countries — ROX™, SOLiD™
Bio-Rad Laboratories, Inc. — CFX384™, CFX96™, Chromo4™, iCycler iQ®, iQ™, MiniOpticon™,
MyiQ™, Opticon™
Cepheid, Inc. — SmartCycler®
Corning, Inc. — Corning®
Diffinity Genomics, Inc. — RapidTip®
Eppendorf AG — Mastercycler®
Excel Scientific, Inc. — AlumaSeal®, SealPlate®, ThermalSeal A™, ThermalSeal RT™, ThermalSeal RT2RR™,
ThermalSeal RTS™, ThermalSeal®
GE Healthcare — Ficoll®
Greiner Bio-One GmbH — SILVERseal™
Illumina, Inc. — Illumina®
Jacobs — Jacobs®
Labnet Intl., Inc. — Multigene™
Life Technologies — SYBR®
Premier Biosoft International — Beacon Designer™
Qiagen GmbH — Rotor-Gene®, Scorpions®
Quanta BioSciences Inc. — KiCqStart®, MystiCq®
Rainin Instrument, LLC — LTS®
Roche Molecular Systems, Inc. — LightCycler®, TaqMan®
Rubicon Genomics, Inc. — GenomePlex®, OmniPlex®, TransPlex®
Sigma-Aldrich Co. LLC — AccuTaq™, DirectLoad™, eAMV™, Extract-N-Amp™, GenElute™, JumpStart™,
LuminoCt®, MTP™, OligoArchitect™, ReadyMix™, ReadyScript®, REDAccuTaq®, REDExtract-N-Amp™,
REDTaq®, Restorase®, SigmaSpin™, SuperPak™
TriLink BioTechnologies, Inc. — CleanAmp™
OligoArchitect™ Online v3.0
Expand your qPCR assay design options with additional
detection chemistries from Sigma® Life Science.
Bioconfident.
In addition to Dual-Labeled Probes and SYBR® Green I Primers,
OligoArchitect Online may now be used to design Molecular
Beacons, Scorpions® Probes, and LightCycler® Probes. Together
with our custom qPCR probes portfolio, Sigma ensures the
success of every qPCR assay — every time.
Design your qPCR assay now
sigma.com/probedesignonline
©2012 Sigma-Aldrich Co. LLC. All rights reserved. SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co. LLC, registered in the US and other
countries. LightCycler is a registered trademark of Roche Molecular Systems, Inc. Scorpions is a registered trademark of Qiagen GmbH. Where bio
begins is a trademark of Sigma-Aldrich Co. LLC.
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Nucleic Acid Amplification

Transcription

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