The application of randomly biotin-labeled BCG small RNA probes by in vitro transcription

Objective Use of biotin-labeled sRNA probes to detect Bacillus Calmette-Guerin (BCG) small RNA (sRNA). Methods The plasmid pMD18-T/ASdes was constructed according to sRNA-ASdes of BCG and used as a template to synthesize the ASdes (3′T7) bio-RNA probes with T7 RNA polymerase by in vitro transcription. Then, the sensibility of the probe was tested with Northern blot and Dot blot. Results The positive signal was obvious in Northern blot in detecting the sRNA-ASdes of BCG using ASdes (3′T7) bio-RNA probes. Conclusion The sensibility of the randomly biotin-labeled sRNA probes obtained by in vitro transcription was high, which could be widely used in the detection of sRNA in Northern blot. Key words: In vitro transcription; Randomly biotin-labeled; sRNA probe; Northern blot; Dot blot