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