Benjak Andrej, Sala Claudia, Hartkoorn Ruben C
Global Health Institute, École polytechnique fédérale de Lausanne (EPFL), CH-1015, Lausanne, Switzerland.
Methods Mol Biol. 2015;1285:17-30. doi: 10.1007/978-1-4939-2450-9_2.
RNA-seq uses next-generation sequencing technology to determine the transcription profile of an organism in a quantitative manner. With respect to microarrays, this methodology allows greater resolution, increased dynamic range, and identification of new features such as previously unannotated genes and noncoding RNAs. Here we describe how to extract RNA from mycobacterial cultures, how to prepare libraries for Illumina sequencing, and the bioinformatics analysis of the sequencing data to determine the transcription profile.
RNA测序利用新一代测序技术以定量方式确定生物体的转录谱。相对于微阵列，这种方法具有更高的分辨率、更大的动态范围，并且能够识别新特征，如先前未注释的基因和非编码RNA。在此，我们描述了如何从分枝杆菌培养物中提取RNA，如何制备用于Illumina测序的文库，以及对测序数据进行生物信息学分析以确定转录谱。
