Waern Karl, Nagalakshmi Ugrappa, Snyder Michael
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT, USA.
Methods Mol Biol. 2011;759:125-32. doi: 10.1007/978-1-61779-173-4_8.
This chapter describes the RNA sequencing (RNA-Seq) protocol, whereby RNA from yeast cells is prepared for sequencing on an Illumina Genome Analyzer. The protocol can easily be altered to use RNA from a different organism. This chapter covers RNA extraction, cDNA synthesis, cDNA fragmentation, and Illumina cDNA library generation and contains some brief remarks on bioinformatic analysis.
本章介绍了RNA测序(RNA-Seq)方案,即如何制备酵母细胞的RNA,以便在Illumina基因组分析仪上进行测序。该方案可轻松修改,以使用来自其他生物体的RNA。本章涵盖RNA提取、cDNA合成、cDNA片段化以及Illumina cDNA文库构建,并对生物信息学分析作了一些简要说明。