Preparation of normalized cDNA libraries for 454 titanium transcriptome sequencing

Zhao Lai, Yi Zou, Nolan C. Kane, Jeong Hyeon Choi, Xinguo Wang, Loren H. Rieseberg

Research output: Chapter in Book/Report/Conference proceedingChapter

9 Scopus citations

Abstract

Transcriptome sequencing from cDNA libraries has been extensively and efficiently used to analyze sequence variation in protein-coding genes (Expressed Sequence Tags) in eukaryote species. Rapid advances in next-generation sequencing (NGS) technology, in terms of cost, speed, and throughput, allow us to address previously unanswerable questions in the fields of ecology, evolution, and systematics using these genomic tools. Transcriptome sequencing from individuals across different populations and species enables researchers to study the evolution of gene sequence variation at a population genomics level. In this chapter, we describe a customized protocol that has been successfully optimized for the development of normalized cDNA libraries in eukaryote systems suitable for Roche 454 GS FLX sequencing, requiring only small quantities of starting material.

Original languageEnglish (US)
Title of host publicationData Production and Analysis in Population Genomics
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages119-133
Number of pages15
ISBN (Print)9781617798696
DOIs
StatePublished - Jan 1 2012
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume888
ISSN (Print)1064-3745

Keywords

  • DSN
  • EST
  • Roche 454 GS FLX
  • Total RNA isolation
  • Transcriptome sequencing
  • cDNA normalization

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Lai, Z., Zou, Y., Kane, N. C., Choi, J. H., Wang, X., & Rieseberg, L. H. (2012). Preparation of normalized cDNA libraries for 454 titanium transcriptome sequencing. In Data Production and Analysis in Population Genomics: Methods and Protocols (pp. 119-133). (Methods in Molecular Biology; Vol. 888). Humana Press Inc.. https://doi.org/10.1007/978-1-61779-870-2_8