RNA Library Preparation

Advances in RNA-Seq library prep are revolutionizing the study of the transcriptome.

Whole Transcriptome

Measure gene and transcript abundance; detect known and novel features in coding and noncoding RNA. Targeted hybridization removes abundant rRNA to focus on high-value portions of the transcriptome.

mRNA

Quantify gene expression, identify known and novel isoforms in the coding transcriptome, detect gene fusions, and measure allele-specific expression.

RNA Enrichment

Analyze gene expression in a focused set of genes of interest. Provides quantitative expression information as well as the detection of small variants and gene fusions.

RNA Amplicon

Use ultra-deep sequencing of polymerase chain reaction (PCR) amplicons for analysis of RNA sequences of interest, differential expression analysis, allele-specific expression measurement, and gene fusion verification.

Time to Compare Custom Enrichment Workflows

Our time-lapse video shows how the entire RNA or DNA workflow can complete before your shift is done, with only 2 hours hands-on time. (DNA workflow shown.)

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Introducing Enhanced RNA-Seq Library Prep Portfolio

These solutions for studying RNA for infectious disease, oncology, and genetic disease research offer rapid turn-around time, broad study flexibility and sequencing scalability.

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Choosing an RNA Kit for Your Experiment

Application	Product	Benefits
Total RNA Sequencing	Illumina Stranded Total RNA Prep 25% off	Integrated enzymatic RNA depletion removes both rRNA and globin mRNA in a single, rapid step One tube to deplete abundant transcripts from multiple species provides flexibility for mixed samples Use with human, mouse, rat, bacteria and epidemiology samples Broad RNA input range, rapid protocol, cost-effective sequencing with up to 384 UDIs Ligation method results in high coverage uniformity and reliability even from degraded samples Measure fold changes in FFPE and low-quality samples
mRNA Sequencing	Illumina Stranded mRNA Prep 25% off	Cost-effective, scalable RNA sequencing of coding transcriptome with precise measurement of strand orientation Broad RNA input range, rapid protocol, cost-effective sequencing with up to 384 UDIs Wide dynamic range for more accurate measurement of gene expression Poly(A) capture, ligation-based addition of adapters and indexes
Targeted RNA Sequencing	Illumina RNA Prep with Enrichment 25% off	Exceptionally fast tagmentation-based library prep enables deep insights into many transcripts of interest, including the RNA exome Single hybridization step No mechanical shearing needed Benefit of panel modularity Compatible with low quality / degraded / FFPE tissue Respiratory virus detection with oligo panel

RNA Library Prep At-a-Glance

Application	Whole transcriptome	mRNA	RNA enrichment
Hands-on time	< 3 hrs	< 3 hrs	< 2 hrs
Turnaround time	~7 hrs	6.5 hrs	< 9 hrs
Input	1 to 1000 ng standard quality RNA; 10 ng for optimal performance and FFPE samples	25 to 1000 ng standard quality RNA	10 ng standard quality RNA; 20 ng RNA for low quality / degraded / FFPE
Automation capability	Liquid handling robots	Liquid handling robots	Liquid handling robots
PCR protocol	No	No	Yes
Library Quant needed?	Yes	Yes	Yes
Fragmentation included?	Yes	Yes	Not required
Product	Illumina Stranded Total RNA Prep	Illumina Stranded mRNA Prep	Illumina RNA Prep with Enrichment

All three kits allow you to decrease sequencing costs by loading up to 384 samples on a single NovaSeq S4 Flow Cell using 384 unique dual indexes for higher throughput sequencing.

The Benefits of Tagmentation

Bead-linked transposome tagmentation is an innovative technology used in our library preparation portfolio. On-bead tagmentation lets you get to sequence-ready libraries faster than before by simultaneously fragmenting the gDNA and adding the Illumina sequencing primers. Normalize your library without ancillary reagents or equipment. Reduce turnaround time and complexity.

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Understanding Adapter Ligation

The other key technology used in our NGS library prep is adapter ligation, long known for consistent, high-quality data. Libraries are prepared by fragmenting a gDNA or cDNA sample and ligating specialized adapters to both fragment ends. These adapters contain the full complement of sequencing primer hybridization sites. This eliminates the need for additional PCR steps, making the process fully automatable.

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Frequently Purchased Together

Ask an expert how you can get the most with Illumina Library Prep

We'll walk through your needs and make recommendations.

RNA-Seq Library Prep Methods and Use Cases

Tale of Two Kits

A critical comparison between two popular RNA Library Prep Kits reveals new information of interest to researchers conducting RNA sequencing studies.

RNA Methods Poster

A comprehensive set of NGS methods for analyzing:

RNA transcription
RNA–protein interactions
Low levels of RNA
RNA modifications
RNA structure

RNA Methods Review

This article collection describes various NGS methods for RNA sequencing, compiled from the scientific literature including pros and cons, publication summaries, and references.

RNA Sequencing Example Workflow for Immuno-oncology Research

Library Prep

llumina RNA Prep with Enrichment

Simultaneously fragment the cDNA and add the Illumina sequencing primers.

Sequencing

NextSeq 2000

Scalable transcriptome and exome sequencing. Process up to 40 samples in a single sequencing run.

Analysis

DRAGEN RNA Pipeline

Perform secondary analysis on the whole flow cell in less than 2 hours.

Product Selection Tools

Library Prep Kit Selector

Find the right sequencing library preparation kit or microarray for your needs. Filter kits by method, species, and more. Compare, share, and order kits.

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Sequencing Method Explorer

Explore library preparation methods for RNA transcription, RNA low-level detection, RNA modifications, RNA structure, and RNA-protein interactions.

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RNA Library Preparation

Simple, Customized RNA-Seq Workflows