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.
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