Next-generation sequencing has transformed gene expression profiling and other RNA expression analysis studies. With an increasing number of researchers employing next-generation sequencing, and with the evolution of sequencing platforms, it is essential to use robust and streamlined methods for sample preparation. The best next-generation sequencing data begin with optimal samples.

Learn more about RNA & Transcriptome Sequencing

RNA Sequencing Overview

Next-generation sequencing studies require robust and streamlined methods for sample preparation. The best next-generation sequencing data begin with optimal samples. Click the tabs above to learn more and see all available products.

RNA Isolation

Although many methods are available for purifying RNA from any particular source, obtaining RNA of the highest purity is the best assurance that the downstream manipulations will be successful. Choose from various formats, including PureLink®, mirVana™, RecoverAll™, and MagMAX™ kits to isolate total RNA, then enrich for the subpopulation of interest.

Simple, reliable, rapid method 30 min isolation from most samples HTP format, ideal for stringent applications Best for next-generation sequencing of mRNA High-throughput purification of FFPE samples
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  PureLink® RNA Mini Kit mirVana™ miRNA Isolation Kit (with or without phenol) MagMAX™ for Microarrays Total RNA Isolation Kit Dynabeads® mRNA DIRECT™ Micro Kit MagMAX™ FFPE Total Nucleic Acid Isolation Kit
RNA types isolated for sequencing Large RNA molecules only (mRNA and rRNA) Small & large RNA molecules (microRNA, tRNA, mRNA, rRNA) Small & large RNA molecules (microRNA, tRNA, mRNA, rRNA) mRNA only total RNA, microRNA, gDNA
Isolation method Fast, convenient silica column Highest purity and convenience; includes organic extraction and silica column Scalable, flexible format for highest purity; includes organic extraction and magnetic beads Fast magnetic bead capture directly from microsized samples Xylene-free, scalable, flexible format with magnetic beads
Prep time 20 min 30 min <1 hr 15 min3 hr (for 96 preps)
Amount of starting material 10 mg to 100 mg of tissue Up to 5 x 107 cells Up to 100 mg of tissue Up to 1 x 107 cells Up to 100 mg of tissue Up to 1 x 107 cells Up to 500 ng purified Total RNA 2 x 10 micron FFPE sections
Prep size 50 preps 40 preps 96 preps 2 mL oligo dT beads (sufficient for 100 mRNA preps) 96 preps
Kit price $225 $225-266 $373 $587$768

RNA Enrichment

To maximize the effectiveness of sequencing reagents, it is often necessary to enrich the sample for specific regions or targets of interest. This enrichment helps eliminate nonspecific artifacts, making it possible to better analyze smaller changes in genomes or transcriptomes and less abundant sequences. Ambion® RiboMinus™ technology is designed to enrich the whole spectrum of RNA transcripts, regardless of their polyadenylation status or the presence of a 5' cap, by selectively depleting rRNA. The RiboMinus™ method removes the vast majority of rRNA (up to 99.9%) to allow greater interrogation of the less abundant transcripts.

Learn more about RiboMinus™ technology

A novel and efficient method utilizing Locked Nucleic Acids (LNA®) to remove up to 99% rRNA A novel and efficient method utilizing Locked Nucleic Acids (LNA®) to remove up to 99% rRNA
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  RiboMinus™ Eukaryote Kit for RNA-Seq RiboMinus™ Plant Kit for RNA-Seq
Platform Diagnostic Diagnostic
Starting material 1–10 µg total RNA 100–500 ng total RNA (low-input protocol) 1–10 µg total RNA
RiboMinus RNA yield 1–2 µg from 10 µg human total RNA ~100 ng from 500 ng human total RNA (low-input protocol) 1–2 µg from 10 µg human total RNA
Probe specificity Eukaryotic Plant
Cat. No. A1083708 - 8 preps A1083808 - 8 preps
Kit price $499.49 $499.49

RNA Library Construction

Library construction, template preparation, and sequencing are at the heart of the sequencing workflow. Utilization of sequencing controls and methods for data analysis complete the workflow and provide useful annotated data. Generate cDNA by reverse transcription from adaptors ligated to the ends of RNA, then amplify the cDNA using primers complementary to the adaptors, and purify it.

Prepare representative cDNA libraries for strand-specific RNA sequencing A Great Solution for NGS The Ultimate Solution for NGS
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  Ion Total RNA-Seq Kit v2 SOLiD® Total RNA-Seq Kit Ambion® RNA-Seq Library Construction Kit
Platform Ion Torrent™ PGM™ SOLiD® 5500 Analyzer Illumina
RNA input type rRNA-depleted total RNA, poly(A) RNA, or miRNA Total RNA, rRNA-depleted total RNA, poly(A) RNA, or miRNA Total RNA, rRNA-depleted total RNA, poly(A) RNA, or miRNA
RNA input range Small RNA: total RNA containing 1–100 ng of miRNA

Whole transcriptome: 100 ng total RNA (~1 ng poly(A) or ~25 ng rRNA-RNA
Small RNA: total RNA containing 1–100 ng of miRNA

Whole transcriptome:
100 ng poly(A), 200 ng
Small RNA: total RNA containing 1–100 ng of miRNA

Whole transcriptome:
100 ng poly(A) or 200 ng rRNA-depleted total RNA, or 200 ng total RNA
Barcodes available Yes 96 No
Cat. No. 4475936 - 12 reaction kit 4445374 - 12 reaction kit 4454073 - 12 reaction kit
Kit price $1,200 $1,433 $1,650

RNA Library Controls


Library construction, template preparation, and sequencing are at the heart of the sequencing workflow. Utilization of sequencing controls and methods for data analysis complete the workflow and provide useful annotated data.

Learn more about ERCC Controls

2 tubes containing preformulated mixes of the same 92 RNA transcripts but in different ratios, for measuring dynamic range, assessing bias, and confirming expression fold changes Single tube containing a preformulated mix of 92 RNA transcripts for measuring dynamic range and assessing bias
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  ERCC ExFold RNA Spike-In Mixes (2 x 10 µL) ERCC RNA Spike-In Mix (1 x 10 µL)
Top Seller
Compatible sample types Eukaryotic total RNA, rRNA-depleted total RNA, or poly(A) RNA Eukaryotic total RNA, rRNA-depleted total RNA, or poly(A) RNA
Dynamic range 6 logs 6 logs
Volume 10 µL (2 tubes) 10 µL
Number of reactions Spike-in controls diluted based on sample type and concentration Spike-in controls diluted based on sample type and concentration
Kit price $2,000 $1,000

RNA Barcoding

Sets of 16 barcodes are available for use with the SOLiD® Total RNA-Seq Kit or the SOLiD® SAGE™ Kit with Barcoding Adaptor Module. Barcodes cost-effectively increase the number of samples that can be analyzed in a single sequencing run. Barcodes assign a unique identifier to templated beads made from a single library. Once assigned, the RNA libraries can be pooled and sequenced.

Cat. No.

Product

4475485

Ion Xpress™ RNA-Seq Barcode 1-16 Kit

4427046

SOLiD® RNA Barcoding Kit, Module 1-16

4453189

SOLiD® RNA Barcoding Kit, Module 17-32

4453191

SOLiD® RNA Barcoding Kit, Module 33-48

4456501

SOLiD® RNA Barcoding Kit, Module 49-64

4456502

SOLiD® RNA Barcoding Kit, Module 65-80

4456503

SOLiD® RNA Barcoding Kit, Module 81-96

4461565

SOLiD® RNA Barcoding Kit, Module 1-48

4461566

SOLiD® RNA Barcoding Kit, Module 49-96

4461567

SOLiD® RNA Barcoding Kit, Module 1-96

RNA Sequencing

No matter which platform you choose to suit your needs, Life Technologies continues to support the evolving needs of this field with gold-standard instrumentation, innovative next-generation technology, optimized best-in-class reagents, consumables, analysis software, applications and world-class technical support.


Next-Generation (SOLiD®)
Sequence right here, right now—minimizing wait time and costs for wasted reagents.
 Semiconductor (Ion Torrent) Sequencing
Sequence from your benchtop with Ion Torrent™ revolutionary "on-chip" technology.
 
Capillary Electrophoresis Sequencing
The gold standard in sequencing and a fast, flexible platform for analysis.
 RNA & Transcriptome Sequencing
Choose the correct technology for your specific RNA application.