RNA Sequencing

Overview

Enables transcriptome profiling, sophisticated gene expression analysis and discovery of new species of RNA.

The transcriptome is comprised of different populations of RNA molecules, including mRNA, rRNA, tRNA, and other non-coding RNA (such as microRNA, lncRNA). RNA sequencing (RNA-seq) using next-generation sequencing (NGS) technology allows us to profile the entire transcriptome, including both the coding and the noncoding regions. It also aids in the identification of genes that are differentially expressed in distinct cell populations and provides information on their relative abundance. In addition, it allows us to determine the effects of genetic variant splicing events, identify novel transcripts, detect gene fusions, isoforms and other structural variants, and call single nucleotide variants (SNVs).

Why RNA Sequencing?

Enables the evaluation of RNA expression levels, variant splicing events, single nucleotide polymorphisms (SNPs), and insertion deletion (indels) events throughout the transcriptome
Provides the flexibility to evaluate specific populations of RNA through different technologies (targeted RNA sequencing, enrichment of mRNA or depletion of rRNA)
Facilitates the detection of low-abundance transcripts and isoforms through targeted sequencing

Workflow

For FFPE RNA

rRNA Depletion
1st and 2nd Strand cDNA Synthesis
RNA Library Preparation
Library Amplification
Library Construction

Workflow

For mRNA

Roche Sequencing Solutions offers products for different parts of this workflow

Workflow Step	Product	Features and Benefits
cDNA Library Preparation	No RNA Enrichment Standard KAPA Stranded RNA-Seq Kits Streamlined KAPA RNA HyperPrep Kits mRNA Capture Standard KAPA Stranded mRNA-Seq Kits Streamlined KAPA mRNA HyperPrep Kits Ribosomal Depletion Standard KAPA Stranded RNA-Seq Kits with RiboErase Streamlined KAPA RNA HyperPrep Kits with RiboErase	RNA-seq or whole transcriptome Traditional or streamlined workflow mRNA-seq Traditional or streamlined workflow Non-coding RNA, whole transcriptome Traditional or streamlined workflow
Library Amplification	KAPA Library Amplification Kit	Minimization of PCR-induced bias of AT- and GC-rich library molecules for improved sequencing coverage of RNA
Library Quantification	KAPA Library Quantification Kits	Accurate quantification of adapter-ligated molecules prior to sample pooling and cluster generation for optimal sequencing results

Workflow Step

Product

Features and Benefits

cDNA Library Preparation

No RNA Enrichment

Standard

KAPA Stranded RNA-Seq Kits

Streamlined

KAPA RNA HyperPrep Kits

mRNA Capture

Standard

KAPA Stranded mRNA-Seq Kits