PrepX RNA-Seq Sample and Library Preparation Kits for Illumina

Transcription

PrepX RNA-Seq Sample and Library Preparation Kits for Illumina
PrepX™ RNA-Seq Sample and
Library Preparation Kits for Illumina
Rapid Total RNA to Library Automation
High-quality stranded libraries from 10 ng of total RNA in a single work day
•Uniform coverage across transcripts down to 10 ng of total RNA
•Streamlined polyA mRNA isolation using the PrepX PolyA mRNA
Isolation Kit or ribosomal depletion using RiboZero*
Consistent and scalable polyA mRNA isolation
•Eliminates the inconsistency and frustration of manual methods
•Higher throughput (48 samples in 2 hours)
•> 95% of rRNA removal
Total RNA to Library
Start a run with as little as 1 sample
•Smart reagent configuration lets you run even 1 sample
with minimal reagent waste
8 samples
48 samples
•Set up each run in ~15 minutes, press RUN, then walk away
~7 hours
~9 hours
* IntegenX’s automated protocol for rRNA works with all of Epicentre’s Ribo-Zero rRNA Removal kits.
Off-the-shelf RNA-Seq Library Automation
The PrepX RNA-Seq sample prepration and library preparation kits on the Apollo 324 System
offer complete walk-away automation of the tedious RNA-Seq library preparation process.
Start with total RNA, and choose between polyA mRNA isolation and rRNA depletion
protocols. The isolated mRNA is ready for QC or can be used directly in the Prep mRNA
library preparation protocols.
PrepX RNA-Seq Sample and Library Preparation Automated Workflow
Total RNA
PolyA mRNA Isolation
rRNA Depletion
PrepX PolyA mRNA Isolation Kit
Ribo-Zero*
mRNA Library Preparation
PrepX RNA-Seq Library Preparation Kit
Fragment
Ligate
Adapter
RT-PCR
PCR (~15 minutes)
Sequence
Bead
Cleanup
B
Manual Isolation
98% Pearson Correlation Between
Apollo 324 and Manual PolyA Isolation
Triplicates of human brain total RNA
Y = 1.1799x - 2.015
R2 = 0.9785
Log RPKM (Apollo 324)
A
no template control
human brain total RNA
human brain total RNA (duplicate)
Apollo 324 Isolation
0% rRNA detected
Log RPKM (Manual)
Figure 2. Gene expression analysis showing 98% Pearson correlation
between the two methods.
Figure 1. Equivalent quality of mRNA isolated from 1 mg of human
brain total RNA (A) manually and (B) on the Apollo 324 System
using the PrepX PolyA mRNA Isolatation Kit.
Figure 3. Uniform coverage of transcriptome for human brain polyA mRNA.
Anti-sense
Sense
Figure 4. Strand specificity of library prep preserved down to 100 pg.
RNA-Seq Sample and Library Preparation Kit Specifications
Protocol
Input
PrepX PolyA PrepX PolyA 48
rRNA Depletion
10 ng to 1 µg of total RNA
up to 1 µg total RNA
PrepX mRNA
PrepX mRNA 48
0.5 to 250 ng polyA mRNA or 200 ng to
1 μg rRNA-depleted mRNA
1 to 8
6 to 48
(multiples of 6)
1 to 8
1 to 8
6 to 48
(multiples of 6)
Setup time
15 minutes
30 minutes
15 minutes
15 minutes
30 minutes
Run time
40 minutes
2 hours
Throughput
(# samples/run)
Kit
Catalog No.
1.5 hours
5 hours
6 hours
PrepX PolyA mRNA
Isolation Kit
(96 samples)
Ribo-Zero rRNA
Removal Kits
(Epicentre)*
PrepX RNA-Seq Library
Preparation Kit for
Illumina (24 samples)
PrepX RNA-Seq Library
Preparation Kit for
Illumina (48 samples)
400047
NA
400039
400046
* IntegenX’s automated protocol for rRNA works with all of Epicentre’s Ribo-Zero rRNA Removal kits.
For more info visit our website:
Toll free: 888.371.4749 • Tel: 925.701.3400 • Fax: 925.701.3318 • www.integenX.com
For research use only. Copyright © 2013, IntegenX Inc. All rights reserved. IntegenX, IntegenX design, Apollo 324 and PrepX are trademarks of IntegenX Inc.
All other trademarks are the sole property of their respective owners. Product specifications may change. D007027 RevA ,3/2013