Lai Zhao, Zou Yi, Kane Nolan C, Choi Jeong-Hyeon, Wang Xinguo, Rieseberg Loren H
The Center for Genomics and Bioinformatics, Indiana University, Bloomington, IN, USA.
Methods Mol Biol. 2012;888:119-33. doi: 10.1007/978-1-61779-870-2_8.
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.
来自cDNA文库的转录组测序已被广泛且有效地用于分析真核生物物种中蛋白质编码基因（表达序列标签）的序列变异。在成本、速度和通量方面，下一代测序（NGS）技术的快速发展使我们能够使用这些基因组工具解决生态学、进化和系统学领域以前无法回答的问题。对不同种群和物种的个体进行转录组测序，使研究人员能够在群体基因组水平上研究基因序列变异的进化。在本章中，我们描述了一种定制方案，该方案已成功优化，用于在真核生物系统中开发适用于罗氏454 GS FLX测序的标准化cDNA文库，只需要少量起始材料。
