Blasdel Bob, Ceyssens Pieter-Jan, Lavigne Rob
KU Leuven, Laboratory for Gene Technology, Kasteelpark Arenberg 21, Box 2642, 3001, Leuven, Belgium.
Bacterial Diseases, Unit Antibiotic Resistance, Scientific Institute of Public Health, Rue Engelandstraat 642, 1180, Brussels, Belgium.
Methods Mol Biol. 2018;1681:185-194. doi: 10.1007/978-1-4939-7343-9_14.
Whole genome wide analysis of transcription using RNA-Seq methods is a powerful way to elucidate differential expression of gene features in bacteria across different conditions as well as for discovering previously exotic RNA species. Indeed, RNA sequencing has revolutionized the study of bacterial transcription with the diversity and quantity of small noncoding RNA elements that have been found and its ability to clearly define operons, promoters , and terminators . We discuss our experience with applying RNA sequencing technology to analyzing the lytic cycle, including extraction, processing, and a guide to the customized statistical analysis necessary for analyzing differential host and phage transcription.
使用RNA测序方法对转录进行全基因组范围的分析，是阐明细菌在不同条件下基因特征差异表达以及发现此前未知的RNA种类的有力手段。事实上，RNA测序已经彻底改变了细菌转录研究，因为已发现的小非编码RNA元件具有多样性和数量众多的特点，并且它能够清晰地界定操纵子、启动子和终止子。我们讨论了将RNA测序技术应用于分析裂解周期的经验，包括提取、处理以及分析宿主和噬菌体转录差异所需的定制统计分析指南。
