Highly consistent, fully representative mRNA-seq libraries from ten nanograms of total RNA

Srikumar Sengupta; Victor Ruotti; Jennifer Bolin; Angela Elwell; Alvaro Hernandez; James A. Thomson; Ron Stewart

doi:10.2144/000113556

Highly consistent, fully representative mRNA-seq libraries from ten nanograms of total RNA

Srikumar Sengupta, Victor Ruotti, Jennifer Bolin, Angela Elwell, Alvaro Hernandez, James A. Thomson, Ron Stewart

Research output: Contribution to journal › Article › peer-review

Abstract

Preparation of an Illumina sequencing library for gene expression analysis (mRNA-Seq) requires microgram amounts of starting total RNA or PCR-based amplification. Here we describe a protocol based on T7 linear RNA amplification that does not introduce significant bias, requires only 10 ng total RNA, and generates a directional, fully representative, whole-transcript mRNA-Seq Illumina library that is highly consistent across over three orders of magnitude of input RNA.

Original language	English (US)
Pages (from-to)	898-904
Number of pages	7
Journal	BioTechniques
Volume	49
Issue number	6
DOIs	https://doi.org/10.2144/000113556
State	Published - Dec 2010
Externally published	Yes

Keywords

Gene expression
High-throughput sequencing
MRNA-seq
Transcriptome

ASJC Scopus subject areas

Biotechnology
General Biochemistry, Genetics and Molecular Biology

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10.2144/000113556

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AU - Stewart, Ron

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Highly consistent, fully representative mRNA-seq libraries from ten nanograms of total RNA

Abstract

Keywords

ASJC Scopus subject areas

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