Literature DB >> 26307248

ARF-TSS: an alternative method for identification of transcription start site in bacteria.

Chao Wang1, Jasmine Lee1, Yinyue Deng1, Fei Tao1, Lian-Hui Zhang1.   

Abstract

Current methods for identifying transcription start sites (TSSs) of specific genes in bacteria usually require adaptors or radioactive labeling. These approaches can be technically demanding and environmentally unfriendly. Here we present a method for identifying TSS called ARF-TSS, which is based on cDNA generation, circularization, PCR amplification, and DNA sequencing to determine the 5'-end of transcripts, thus circumventing the need for adaptors and radioactive labeling. We validated the method using the gene lasI from the bacterial pathogen Pseudomonas aeruginosa. Our results show that ARF-TSS could be a good alternative to traditional methods for bacterial TSS analysis.

Entities:  

Keywords:  Pseudomonas aeruginosa; mRNA; promoter; reverse transcription; transcriptional initiation; transcripts

Mesh:

Substances:

Year:  2012        PMID: 26307248     DOI: 10.2144/000113858

Source DB:  PubMed          Journal:  Biotechniques        ISSN: 0736-6205            Impact factor:   1.993


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