Singh Navjot, Wade Joseph T
Wadsworth Center, New York State Department of Health, Albany, NY, USA.
Methods Mol Biol. 2014;1103:1-10. doi: 10.1007/978-1-62703-730-3_1.
The ability to map transcription start sites is critical for studies of gene regulation and for identification of novel RNAs. Conventional RNA-seq is often insufficient for identification of transcription start sites due to low coverage and/or RNA processing events. We have developed a highly sensitive, genome-scale method for detection of transcription start sites in bacteria. This method uses deep sequencing of cDNA libraries to identify transcription start sites with strand specificity at nucleotide resolution. Here, we describe the application of this method for transcription start site identification in Escherichia coli.
绘制转录起始位点的能力对于基因调控研究和新型RNA的鉴定至关重要。由于覆盖度低和/或RNA加工事件,传统的RNA测序通常不足以鉴定转录起始位点。我们开发了一种高度灵敏的、全基因组规模的方法来检测细菌中的转录起始位点。该方法利用cDNA文库的深度测序,以核苷酸分辨率在链特异性水平上鉴定转录起始位点。在此,我们描述了该方法在大肠杆菌中转录起始位点鉴定中的应用。