Shell Scarlet S, Chase Michael R, Ioerger Thomas R, Fortune Sarah M
Department of Immunology and Infectious Diseases, Harvard School of Public Health, 677 Huntington Avenue, Boston, MA, 20115, USA.
Methods Mol Biol. 2015;1285:31-45. doi: 10.1007/978-1-4939-2450-9_3.
Next-generation sequencing technologies facilitate the analysis of multiple important properties of the transcriptome in addition to gene expression levels. Here we describe a method for mapping RNA 5' ends in Mycobacterium tuberculosis, which allows the determination of transcriptional start sites (TSSs), comparative analysis of promoter usage under different conditions, and mapping of endoribonucleolytic processing sites. We describe in detail the procedures for constructing RNA sequencing libraries appropriate for RNA 5' end mapping using an Illumina sequencing platform. We also outline the major steps of data analysis.
除了基因表达水平外，新一代测序技术还便于对转录组的多个重要特性进行分析。在此，我们描述了一种在结核分枝杆菌中绘制RNA 5'末端图谱的方法，该方法可用于确定转录起始位点（TSS）、不同条件下启动子使用情况的比较分析以及核糖核酸内切酶加工位点的图谱绘制。我们详细描述了使用Illumina测序平台构建适用于RNA 5'末端图谱绘制的RNA测序文库的步骤。我们还概述了数据分析的主要步骤。
