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RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...

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Updated: May 11, 2026

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA

Published on: October 27, 2011

Tips for preparing mRNA-Seq libraries from poly(A)+ mRNA for Illumina transcriptome high-throughput sequencing.

Brenton R Graveley

Cold Spring Harbor Protocols

|May 3, 2013

Summary

This guide offers background, tips, and troubleshooting for Illumina's mRNA sequencing (mRNA-Seq) library preparation kits. It aims to assist researchers using these continually updated tools for high-throughput gene expression analysis.

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Last Updated: May 11, 2026

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA

Published on: October 27, 2011

AQRNA-seq for Quantifying Small RNAs

AQRNA-seq for Quantifying Small RNAs

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Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs

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Published on: September 16, 2019

Area of Science:

Molecular Biology
Genomics
Bioinformatics

Background:

High-throughput sequencing of messenger RNA (mRNA-Seq) is a key technique in gene expression analysis.
Researchers commonly utilize commercial kits for mRNA library preparation.
Illumina kits are widely adopted but subject to frequent updates.

Purpose of the Study:

To provide essential background information on Illumina mRNA library preparation kits.
To offer practical tips for optimizing library preparation workflows.
To present troubleshooting advice for common issues encountered with these kits.

Main Methods:

Review of current Illumina mRNA library preparation kits.
Compilation of best practices and experimental tips.
Identification and explanation of frequent technical challenges and solutions.

Main Results:

Detailed overview of available Illumina mRNA library preparation kits.
Actionable recommendations for successful library construction.
Guidance on resolving common problems in mRNA-Seq library preparation.

Conclusions:

Understanding and applying these tips can improve mRNA-Seq data quality.
Effective troubleshooting is crucial for reliable high-throughput sequencing results.
This resource aids researchers in maximizing the utility of Illumina's evolving mRNA library preparation kits.