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Methods Mol Biol. 2013;923:221-39.

Transcriptome analysis using RNA-Seq.

Author information

1
Department of Molecular Biology, Nijmegen Center for Molecular Life Sciences, Nijmegen, The Netherlands.

Abstract

Transcriptome analysis by next-generation sequencing (RNA-seq) allows investigation of a transcriptome at unsurpassed resolution. One major benefit is that RNA-seq is independent of a priori knowledge on the sequence under investigation, thereby also allowing analysis of poorly characterized Plasmodium species. Here we provide a detailed protocol for RNA isolation and fragmentation, ribosomal RNA depletion, and cDNA synthesis that enables the preparation of a sequencing library from 1 to 2 μg of total RNA. Although we focus our discussion on the quantitative measurement of gene expression, this protocol is suited for many applications of RNA-seq and allows analysis of most RNA species.

PMID:
22990781
DOI:
10.1007/978-1-62703-026-7_15
[Indexed for MEDLINE]

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