nucleic acid purification kits - Omega Bio-tek

NUCLEIC ACID PURIFICATION KITS
FAST
SIMPLE
QUALITY
CONVENIENT
REPRODUCIBLE
Innovations in Nucleic Acid Isolation
COMPANY PROFILE
Since its founding in 1998, Omega Bio-tek, Inc. has been at the forefront of nucleic acid purification by offering
products for clinical and basic research, biotechnology and agricultural applications. DNA and RNA extraction is the first
step for so many downstream analyses, and efficient and clean nucleic acid isolation is crucial. Our goal is to offer high
quality products to help you improve your workflows. Our diverse product line ranges from RNA purification from
plants to DNA extraction from dried blood spots. By
offering over 900 products for nucleic acid isolation,
we have a solution to almost any of your nucleic acid
purification needs. Contact one of our specialists to
see which product would best fit your application.
With the ability to offer individual components and
customize kits, we can help your lab reduce waste and
increase productivity.
Quality is key to operations at Omega Bio-tek. We
offer products for manual and automated
processing and we support our customers by having
Hamilton STAR®, Beckman Coulter Biomek® FX and
Thermo KingFisher® instruments in house. In addition
to offering nucleic acid purification products, we also
offer next generation sequencing services utilizing
Illumina platforms. We are ISO 9001:2008 certified and
we ensure that our products are properly assembled, tested, recorded, stored and shipped. We perform rigorous quality
checks of our products and thoroughly train our employees to ensure compliance. We also have several quality control
steps within our processes to deliver the best product. We firmly believe that quality in equals quality out.
THE OMEGA DIFFERENCE
Our diverse portfolio consists of 900+ products ranging from RNA isolation from plants to DNA extraction from dried
blood spots. With the ability to offer individual components separately and to customize kits, we can help your lab reduce waste and increase productivity.
•
ECONOMICAL:
On average, Omega Bio-tek’s
products cost 30% less than the competition.
•
CUSTOMIZABLE:
For large customers, we can generate custom
packaging, kits, reagents, etc.
•
INDIVIDUAL COMPONENTS:
We sell kit components separately.
•
PRODUCT RANGE:
Alternative options for almost all of your nucleic acid
purification needs.
01
E.Z.N.A.® PLASMID MINI KIT I
The E.Z.N.A.® Plasmid Mini Kit I is designed to isolate up to 30 µg of high quality plasmid DNA from 1-5 mL
bacterial cultures in 30 minutes or less. This kit uses a modified alkaline lysis method to lyse the cells and
separate genomic DNA from plasmid DNA. Plasmid DNA purification is further simplified by using our
HiBind mini column technology in three quick steps: bind, wash and elute. Purified plasmid DNA is ready for
a wide variety of downstream applications, including routine screening, restriction enzyme digestion, DNA
sequencing, cloning, transformation and transfection. The E.Z.N.A.® Plasmid Mini Kit I is available in two
different column formats: V-spin mini columns (with an attached cap) and Q-spin mini columns (without a
cap).
PLASMID KIT PROTOCOL
Pellet cells by centrifugation
DNA CONCENTRATION COMPARISON
Omega Bio-tek
Resuspend
Lyse
Neutralize
Transfer cleared
lysate to HiBind® Mini
Column
Plasmid DNA in ng/µL
Clear Lysate
Company Q
300
250
200
150
100
50
0
Bind
DNA Concentration Comparison. 4 mL DH5*
cultures transformed with pGEM vector were
performed
according
to
manufacturer’s
recommended
protocols.
Plasmid
DNA
concentration determined by optical density
measurements with NanoDrop 2000c. Total elution
volume used was 50 µL.
Product No.
Wash 3x
Dry
Elute
Preps
Q-spin (no cap)
D6942-00
5
D6942-01
50
D6942-02
200
V-spin (cap attached)
D6943-00
5
D6943-01
50
D6943-02
200
02
FASTFILTER PLASMID DNA MIDI & MAXI KITS
The FastFilter Plasmid Kits combine the power of HiBind technology with the time-tested consistency of
alkaline-SDS lysis of bacterial cells to deliver high quality plasmid DNA. This system includes a special lysate
clearance syringe, which replaces the centrifugation step following alkaline lysis. Plasmid DNA is suitable for
many downstream applications including sequencing, restriction digestion and transfection.
FASTFILTER PLASMID DNA KITS
PROTOCOL
PROCESSING TIME OF
PLASMID DNA ISOLATION KITS
Pellet bacterial cells by centrifugation
Resuspend
Lyse
150
120
Neutralize
Add Binding Buffer
90
Transfer lysate into Lysate Clearance
Syringe
60
30
Insert column into luer connector on
vacuum manifold
Pass lysate through DNA column
Wash 3x
Centrifuge to dry
Elute by centrifugation
03
ny
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Co
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ny
Q
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ny
Q
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Co
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io
aB
eg
Om
Prepare vacuum manifold
HS
0
Insert plunger into Lysate Clearance
Syringe and clear lysate
Processing time of plasmid DNA
isolation kits. Total time in minutes
required for plasmid DNA isolation
according to the manufacturer’s
recommended protocols for the
Omega Bio-tek FastFilter Plasmid
DNA Kit and three products from
competitors.
Product No.
Preps
FastFilter Plasmid DNA Mini Kit
D6905-01
5
D6905-03
25
D6905-04
100
FastFilter Plasmid DNA Maxi Kit
D6924-01
5
D6924-03
25
D6924-04
100
E.Z.N.A.® PLASMID MIDI & MAXI KIT
The E.Z.N.A.® Plasmid Midi Kit can isolate up to 200 µg from a 50 mL bacterial culture, and the E.Z.N.A.® Plasmid
Maxi Kit can isolate up to 1 mg from a 200 mL culture. This kit uses a modified alkaline lysis method to lyse
the cells and separate genomic DNA from plasmid DNA. Cellular debris is removed by centrifugation, and the
cleared lysate is applied to a HiBind DNA column, which eliminates time consuming steps such as isopropanol
precipitation. Plasmid DNA is suitable for automated fluorescent DNA sequencing, restriction endonuclease
digestion, transfection of mammalian cells and many other downstream applications.
PLASMID DNA YIELDS FROM E.Z.N.A.® KITS
E.Z.N.A.® Plasmid DNA Midi Kit
Sample
1
2
3
4
Culture Size
50 mL
50 mL
50 mL
50 mL
A260/A280
1.89
1.90
1.90
1.90
A260/A230
2.31
2.40
2.39
2.39
Yield (µg)
192.9
189.0
190.0
187.4
E.Z.N.A.® Plasmid DNA Maxi Kit
1
2
3
4
250 mL
250 mL
250 mL
250 mL
1.90
1.90
1.90
1.90
2.29
2.34
2.35
2.32
715.2
697.0
706.5
701.3
Plasmid DNA Yields from E.Z.N.A.® Kits. DH5* cells were transformed with
pGEM vector, and replicate bacterial cultures were grown in either 50 mL or
250 mL of LB broth for 24 hours. The E.Z.N.A.® Plasmid Midi Kit was used to
isolate plasmid DNA from the 50 mL cultures and the E.Z.N.A.® Plasmid Maxi
Kit was used to isolate plasmid DNA from the 250 mL cultures. Yield was
determined by optical density measurements with the NanoDrop 2000c.
Product No.
Preps
E.Z.N.A.® Plasmid DNA Midi Kit
D6904-00
2
D6904-03
25
D6904-04
100
E.Z.N.A.® Plasmid DNA Maxi Kit
D6922-01
5
D6922-02
25
D6922-04
100
04
E.Z.N.A.® CYCLE PURE KIT
The E.Z.N.A.® Cycle Pure Kit is designed for the rapid purification of single- or double-stranded DNA
from PCR or other enzymatic reactions. The purification procedure completely removes primers,
nucleotide enzymes, salts and other impurities from the DNA sample. The DNA sample is simply mixed
with buffer and spun through the HiBind DNA column. The DNA bound to the HiBind matrix is washed
and the clean, concentrated DNA is eluted with deionized water or elution buffer. This convenient spin
column format eliminates the need for expensive resins or toxic organic compounds such as phenol and
chloroform, thereby making it possible to process multiple samples in parallel. Purified DNA can be
used in T-A ligations, sequencing, restriction enzyme digestions and various other labeling reactions.
any Q
Com
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Ome y P
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Com
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T
Com
pany
A
Com
pany
Q
Com
pany
P
Ome
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any A
Com
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Com
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M
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any T
OMEGA BIO-TEK’S CYCLE PURE KIT
VS. THE COMPETITION
CYCLE PURE PROTOCOL
Add Buffer CP to PCR
sample
M
Apply CP/PCR to a HiBind™
Column
Wash with DNA
Wash Buffer
PCR products in two sizes (500 bp and 5 kb) were
purified with four different competitor’s kits and
Omega Bio-tek’s E.Z.N.A.® Cycle Pure Kit. Ten
percent of eluted product was analyzed on a 0.8%
agarose gel and run with a DL2000 marker.
Dry
Elute
05
Product No.
Preps
D6492-00
D6492-01
D6492-02
5
50
200
E.Z.N.A.® GEL EXTRACTION KIT
Omega Bio-tek’s E.Z.N.A.® Gel Extraction Kit uses HiBind DNA mini column technology to recover DNA bands
from 100 bp to 20 kb in size. This kit can be used with all concentrations and types of agarose gels, including
TBE and TAE gels with a maximum recovery rate of 85%.
The DNA band of interest is excised from the gel, dissolved in binding buffer and applied to a DNA HiBind
mini column. Following three wash steps, DNA is eluted with elution buffer and is ready for downstream
applications such as ligation, PCR amplification, restriction enzyme digestion and various labeling reactions.
The E.Z.N.A.® Gel Extraction Kits can also be used to purify DNA fragments from PCR products and other
enzymatic reactions.
RECOVERY RATE OF EXCISED DNA
VS COMPANY Q
Control
Omega Bio-tek
Recovery Rate %
Company Q
100
80
Product No.
Preps
D2500-00
D2500-01
D2500-02
5
50
200
60
40
20
5000 bp
500 bp
1000 bp
M
C
O
Q
C
O
Q
C
O
200 bp
Q
C
O
Q
M
Percent recovery of 4 different sizes of DNA bands
from a 2% agarose gel with the E.Z.N.A.® Gel
Extraction Kit (O, green) and a comparable
kit from Company Q (Q, blue) according to
manufacturer’s recommended protocols. The
original input amounts of DNA (C, black) were
normalized to 100% and the amount of DNA
recovered was determined by optical density
measurements with the NanoDrop 2000c.
06
E.Z.N.A.® TISSUE DNA KIT
The E.Z.N.A.® Tissue DNA Kit offers a simple, rapid and cost effective method for the isolation of DNA from a
wide variety of sample sources including fresh and frozen tissues, animal cells, buccal swabs and FFPE tissue
samples. After cell lysis, the DNA purification process can be completed in less than 30 minutes. Up to 30 mg
of tissue can be used, and multiple samples can be simultaneously processed with this spin column-based
kit. There is no need for time consuming steps such as phenol/chloroform extractions or isopropanol/ethanol
precipitations. DNA purified using the E.Z.N.A.® Tissue DNA Kit is ready for most downstream applications
such as PCR, Southern blotting, microarrays, SYBR or probe-based qPCR and restriction enzyme digestion.
Sample
Sample Size
Whole blood
Buffy coat
Cultured cells
Buccal swabs
Saliva
200 µL
200 µL
1 x 106
1 swab
200 µL
REAL-TIME PCR OF GENOMIC DNA
ISOLATED WITH E.Z.N.A.® TISSUE DNA
KIT
Range of Yield
(µg)
4-8
30-40
5-6
2-4
6-10
Genomic
wasfrom
isolated
from
10 mg
ratE.Z.N.A®
kidney
DNA wasDNA
isolated
10mg rat
kidney
usingofthe
with
Omega
Bio-tek’s
E.Z.N.A.®
Tissue
DNA
Kit.
SeTissue DNA Kit . A serial dilution of recovered DNA was then
rial dilutions of recovered genomic DNA were used
used in a SYBR Green real-time qRT-PCR targeting a 100 bp
as templates
for real-time PCR amplification of a 100
of the
gene. Each
reaction
performed
bp fragment
fragment
of GAPDH
the GAPDH
gene
with was
SYBR
Green in
3 replicates.
lines represent
input DNA amounts
of 10, 2,
labeling.
EachThe
reaction
was performed
in triplicate.
0.4,
0.08,
and
0.016ng
(left
to
right)
per
reaction.
The fluorescence versus cycle number is plotted
above and the five curves correspond to the input
DNA template amounts of 10, 2, 0.4, 0.08 and 0.0016
ng.
YIELD COMPARISON OF OMEGA
BIO-TEK’S Rat
TISSUE
DNA KIT
kidney
M
M
T
Company T
A
Company A
P
Company P
Omega
Q
Company Q
Purified genomic DNA, from fresh rat tissue, was isolated with the company T、
company
A、company
P、E.Z.N.A.®
Tissuefrom
DNA Kit、company
Tissuekidney
DNA
Purified
genomic
DNA
10 mg Qrat
Kit. DNA was extracted from 10mg kidney tissue samples of rat. Genomic DNA
tissue was isolated with Company T, Company A,
(3% of total purified DNA) was analyzed on a 0.8% agarose gel to demonstrate
Company P, Omega Bio-tek’s Tissue DNA Kit and
yield and quality of the DNA.
Company Q’s genomic DNA extraction kits following
M:lambda-HindIII
the manufacturer’s recommended protocols. Three
percent of eluted DNA was analyzed on a 0.8%
agarose gel. M: Lambda-Hind III.
07
M
Product No.
Preps
D3396-00
D3396-01
D3396-02
5
50
200
E.Z.N.A.® TOTAL RNA KIT I
Source
Brain
Kidney
Liver
Heart
Spleen
Lung
Pancreas
Thymus
IC-21
HeLa
HEK-293
HIN3T3
Sample Size
10 mg
10 mg
10 mg
10 mg
10 mg
10 mg
10 mg
10 mg
1 x 106 cells
1 x 106 cells
1 x 106 cells
1 x 106 cells
RNA Yield (µg)
10
30
45
5
33
12
40
20
12
15
10
15
Expected RNA yields from mouse tissue samples and
cultured cell types with the E.Z.N.A.® Total RNA Kit 1.
TOTAL RNA EXTRACTION QUALITY
WITH THE E.Z.N.A.® TOTAL RNA KIT I
1
2
3
4 5
6 7
8
9
TOTAL RNA EXTRACTION
CONCENTRATIONS WITH THE
E.Z.N.A.® TOTAL RNA KIT I
350
300
ng/µL
The E.Z.N.A.® Total RNA Kit I provides a simple and
rapid method for the isolation of up to 100 µg
total RNA from cultured eukaryotic cells and soft
tissues. This kit enables simultaneous processing of
multiple samples in less than 20 minutes. Typically, up
to 1 x 107 eukaryotic cells or 30 mg tissue can be used
in a single reaction. Many different sample types can
be used including kidney, liver, heart, spleen, lung,
pancreas, thymus, HeLa cells, HEK-293 cells and
HIN3T3 cells. Purified RNA can be used in many
downstream applications such as RT-PCR, qRTPCR, Northern blotting, nuclease protection assay,
microarrays, in vitro translation and next generation
sequencing.
250
200
150
100
50
0
Product No.
Preps
R6834-00
R6834-01
R6834-02
5
50
200
1
2
3
4
5
6
7
8
9
Samples
Total RNA was isolated from 2.5 106 HEK-293 cultured
cells with Omega Bio-tek’s E.Z.N.A.® Total RNA Kit I
with an elution volume of 50 µL. RNA concentrations
were quantified with Thermo NanoDrop 2000C. RNA
quality was assessed with the Agilent Tapestation
2200.
08
E.Z.N.A.® PLANT DNA DS KIT
The E.Z.N.A.® Plant DNA DS Kit is designed for the efficient recovery of genomic DNA up to 30 kb in size from
fresh, frozen or dried plant tissue samples rich in polysaccharides, polyphenols or having a lower DNA content.
Up to 50 mg of wet tissue can be processed in less than 1 hour. The system combines the reversible nucleic
acid-binding properties of the HiBind® matrix with the speed and versatility of spin column technology
to eliminate polysaccharides, phenolic compounds and enzyme inhibitors from plant tissue lysates. Purified
DNA is suitable for PCR, restriction digestion and hybridization applications.
This procedure relies on the well-established properties of the cationic detergent cetyltrimethyl ammonium
bromide (CTAB) in conjunction with the unique binding system to increase yields and provide high quality
DNA. The system eliminates the need for chloroform extractions traditionally associated with CTAB-based
lysis methods. Samples are homogenized and lysed in a high salt buffer containing CTAB, binding conditions
are adjusted and DNA is purified using a HiBind® DNA mini column. Salts, proteins and other contaminants
are removed to yield high quality genomic DNA suitable for downstream applications such as endonuclease
digestion, thermal cycle amplification and hybridization applications.
COMPARISON OF DNA YIELD
FROM MULTIPLE CROPS
150
Omega Bio-tek
Company Q
120
Product No.
60
30
ac
co
To
b
n
ea
So
yb
to
n
Co
t
n
Co
r
la
0
Ca
no
ng/mg
90
40-50 mg of respective fresh leaf tissue were extracted
in triplicate according to the manufacturer’s
recommended protocol and eluted in 100 µL. DNA
analyzed with flourescent DNA-based quantification
method. Total yield was divided by total tissue
amount to show ng of DNA per mg of leaf tissue.
09
Preps
E.Z.N.A.® Plant DNA DS Kit
D2411-00
5
D2411-01
50
E-Z 96® Plant DNA DS Kit
D1411-00
1 x 96
D1411-01
4 x 96
E.Z.N.A.® VIRAL RNA KIT
The E.Z.N.A.® Viral RNA Kit is designed for the
isolation of viral RNA from acellular fluids such as
plasma, serum and urine. The kit can also be used
to isolate viral DNA, genomic DNA, circulating DNA
and total RNA. The procedure completely removes
contaminants and enzyme inhibitors making
viral RNA isolation fast, convenient and reliable. RNA
purified using the E.Z.N.A.® Viral RNA method is ready
for all downstream applications such as RT-PCR and
qPCR. This kit has been validated for isolating viral
nucleic acids from hepatitis A, B, C and HIV.
RNA Template
1 x 107 viral particles/µL
1 x 106 viral particles/µL
1 x 105 viral particles/µL
1 x 104 viral particles/µL
REAL-TIME PCR OF VIRAL RNA
ISOLATED WITH E.Z.N.A.® VIRAL RNA
KIT
1 x 103 viral particles/µL
cT Value
19.90
19.88
19.98
23.09
23.09
22.99
25.48
25.53
25.08
28.64
28.56
28.66
31.23
31.59
31.58
Amplification
3000
RFU
2000
1000
0
0
10
20
30
40
Cycles
Serum was separated from a human blood sample
containing 1 x 107 Hepatitis B viral particles/µl, a 10fold dilution series of the serum was performed and
50 µl of each dilution was used in the Mag-Bind®
Viral RNA/DNA Kit to isolate viral RNA. Two microliters
of recovered RNA from each dilution of serum was
used as a template in a real-time PCR reaction with
SYBR Green labeling. Each reaction was performed in
triplicate. The 5 curves represent the fluorescence
versus cycle number for the 5 starting serum
concentrations.
Product No.
Preps
R6874-00
R6874-01
R6874-02
5
50
200
10
400 Pinnacle Way Suite 450
Norcross, GA 30071
Phone: (800) 832-8896 / Fax: (770) 931-0230
[email protected]
www.omegabiotek.com
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