Ion Total RNA-Seq Kit v2

The Ion Total RNA-Seq Kit v2 contains everything needed to construct representative cDNA libraries for strand-specific RNA sequencing on the Ion PGM™ Sequencer. Utilizing proprietary technology that inhibits synthesis of a reaction by-product, the gel size selection step has been eliminated from the small-RNA library workflow. Researchers can now prepare libraries to analyze small RNA, including microRNA, or perform gene expression profiling in just 6 hours with an easy-to-use, automation-friendly, magnetic bead–based cleanup. This kit is an integral part of the fast, scalable, and affordable Ion RNA-Seq solution, ideal for researchers seeking an alternative to microarrays that is more sensitive in detecting transcripts and changes in expression and that offers the ability to discover novel and fusion transcripts and identify isoforms.

* Typically >500,000 reads mapped to miRBase per library made from human placenta RNA ** Typically >2 million reads mapped to RefSeq exons with a HeLa whole-transcriptome library using

poly(A) RNA as the starting material

Ion Total RNA-Seq Kit v2

Library type Small RNA, including microRNA Whole transcriptome

Input RNA required As little as:

• 5 ng miRNA in 100 ng of total RNA

• 1 ng miRNA in RNA enriched for small RNA

As little as: • 1 ng of poly(A) RNA isolated

from 100 ng total RNA • 25 ng of rRNA-depleted RNA

isolated from 100 ng total RNA

Time ~6 hours

Strand information preserved Yes

Cleanup technology Magnetic bead–based cleanup, no gel size selection or filter columns

Barcode-compatible Yes, Ion Xpress™ RNA-Seq Barcode 1–16 Kit for multiplexing

Ion 318™ Chip capacity 3 barcoded small-RNA libraries* 1 whole-transcriptome library**

Discover what you have been missing at lifetechnologies.com/ionrnaseq

See the clear picture with Ion RNA-Seq and reveal the diversity of the transcriptome

Survey the small-RNA or mRNA transcriptome Construct libraries for the analysis of small RNA, including microRNA, or perform gene expression profiling by sequencing.

Accelerated workflow Construct libraries in just 6 hours without labor-intensive gel size selection for small-RNA libraries, and achieve effective, rapid cleanup with automation-friendly magnetic beads.

Visit the Ion Community to view:

• Application notes covering sequencing for small-RNA analysis and gene expression profiling

• Videos

• How-to guides

• Data sets

Figure 1. Ion RNA-Seq workflow. Times given are based on a workflow using an Ion 318™ Chip and barcoded small-RNA libraries. Times for library construction and template preparation for the workflow for gene expression by sequencing are the same; however, sequencing and data analysis for the longer 100-base run are 2.5 hours and 2 hours, respectively.

For research use only. Not intended for any animal or human therapeutic or diagnostic use. ©2012 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. CO25394 0512

Preserve strand information Proprietary Ambion® technology attaches adapters in a directional manner that preserves strand information in the libraries. As a result, mapped reads are aligned in the direction of transcription relative to the chromosomal strand, which aids in the understanding of the structure and expression levels of the transcript and in the discovery of novel transcription regions.

Optimize performance with RNA isolation products The new “mRNA from total RNA” protocol for the Dynabeads® mRNA DIRECT™ Micro Kit has been optimized to further reduce rRNA content. When poly(A) RNA purified with this kit and protocol are used to make whole-transcriptome libraries using the Ion Total RNA-Seq Kit v2, we observe a 10-fold reduction in the number of reads mapping to rRNA and a corresponding increase in reads mapping to RefSeq exons. Use the mirVana™ miRNA Isolation Kit to isolate small RNA from tissues and cells for the construction of small-RNA libraries using the Ion Total RNA-Seq Kit v2.

Tailor read number and read length to your needs Ion 314™, 316™, and 318™ Chips and 16 RNA-Seq barcodes enable researchers to scale the number of reads for the application. In addition, researchers can define the read length of the Ion PGM™ Sequencer run (35, 100, or 200 bases) to best suit their needs.

Move rapidly to results Construct libraries in just 6 hours using the Ion Total RNA-Seq Kit v2 and Ion Xpress™ RNA-Seq Barcodes (Figure 1). Template is prepared from the libraries using the Ion OneTouch™ System and sequenced on the Ion PGM™ Sequencer. Data analysis is performed with Torrent Suite Software, and outputs are provided in standard file formats that can be used by many analysis programs.

Ordering information

Product Cat. No.

Ion Total RNA-Seq Kit v2 4475936

Related products

Ion Xpress™ RNA-Seq Barcode 1-16 Kit 4475485

mirVana™ miRNA Isolation Kit AM1560

Dynabeads® mRNA DIRECT™ Micro Kit 61021

ERCC RNA Spike-In Mix 4456740

CONSTRUCTLIBRARY

ION TOTAL RNA-SEQ KIT v2 ION ONETOUCH™ SYSTEM ™ION PGM SEQUENCER TORRENT SUITE

PREPARETEMPLATE

RUN SEQUENCE

ANALYZE DATA

4HOURS

2HOURS

1HOUR

6HOURS