Chapter title |
High-throughput RNA sequencing in B-cell lymphomas.
|
---|---|
Chapter number | 17 |
Book title |
Lymphoma
|
Published in |
Methods in molecular biology, January 2013
|
DOI | 10.1007/978-1-62703-269-8_17 |
Pubmed ID | |
Book ISBNs |
978-1-62703-268-1, 978-1-62703-269-8
|
Authors |
Xiao W, Tran B, Staudt LM, Schmitz R, Wenming Xiao, Bao Tran, Louis M. Staudt, Roland Schmitz |
Abstract |
High-throughput mRNA sequencing (RNA-seq) uses massively parallel sequencing to allow an unbiased analysis of both genome-wide transcription levels and mutation status of a tumor. In the RNA-seq method, complementary DNA (cDNA) is used to generate short sequence reads by immobilizing millions of amplified DNA fragments onto a solid surface and performing the sequence reaction. The resulting sequences are aligned to a reference genome or transcript database to create a comprehensive description of the analyzed transcriptome. This chapter describes a protocol to perform RNA-seq using the Illumina sequencing platform, presents sequencing data quality metrics and outlines a bioinformatic pipeline for sequence alignment, digital gene expression, and mutation discovery. |
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