RNA Sequencing

Elucidating changes in gene expression is fundamental to understanding cellular response to internal genetic lesions, external stimuli, and changing environmental conditions. As a consequence, RNA expression patterns present a unique source of biomarkers for prediction and classification of human disease. While much focus to date has been placed on protein encoding RNAs (so called poly-A+ RNA), the importance of non-coding RNA sequences is well documented. Next-generation RNA sequencing is a highly compelling platform for RNA research, offering the following key benefits:

  • ‘Digital counting’ of RNA molecules for highly quantitative and precise measurements
  • Dynamic range to fully capture relevant biological changes
  • Discovery of unknown RNAs (novel transcripts, splice variants, and gene fusions)
  • Capture multiple RNA types (poly-A+, long non-coding RNA, and gene fusions) in a single assay
  • Opportunity to focus from complete whole transcriptome analysis down to a handful of pre-selected RNA sequences to balance experimental cost and ease of analysis with discovery potential

RNA sequencing with Ion Torrent NGS technology

From targeted gene expression analysis to whole transcriptome sequencing, Ion Torrent™ products combine simple sample preparation and intuitive data analysis to deliver a comprehensive RNA sequencing solution.

RNA Sequencing opposites

Targeted transcriptome sequencing
A targeted sequencing approach focusing on >20,000 known genes—a great compromise to whole transcriptome sequencing as analysis if vastly simplified

Whole transcriptome sequencing
Also referred to as RNAseq, this  approach has the greatest discovery potential but, as a consequence, poses the greatest bioinformatics challenges

Strand-specific, hypothesis-free whole transcriptome analysis enables identification and quantification of both known and novel transcripts. Ion Torrent next-generation sequencing systems, combined with Invitrogen RNA purification and Ion Torrent library construction kits, offer a reliable sequencing workflow that combines simple sample preparation and intuitive data analysis for your whole transcriptome sequencing.

Whole transcriptome sequencing is great for discovery, but overly complex for traditional gene expression studies. With a greater quantitative linear dynamic range compared to microarray technology, take advantage of the speed and increased sensitivity of next-generation sequencing (NGS) while keeping sample preparation and data analysis simple with the Ion AmpliSeq™ Transcriptome Human Gene Expression Kit. Target >20,000 RefSeq transcripts using as little as 10 ng RNA input with fast and formalin-fixed, paraffin-embedded (FFPE)-compatible sample preparation, sequencing, and data analysis workflows for simple gene-level expression analysis.

Targeted RNA sequencing is a highly affordable solution for following up on transcriptome studies, or it can be used to rapidly focus in on pathway- or disease-related content. With very low amounts of input total RNA (as low as 500 pg of unfixed RNA or 5 ng of RNA isolated from FFPE samples), expression of a handful, hundreds or even thousands of RNA targets can be assayed in a simple NGS-based workflow enabling the monitoring of messenger RNA (mRNA), non-coding RNA (ncRNA), and gene fusions.

For Research Use Only. Not for use in diagnostic procedures.