Application of Individual Bacterial Colonies PCR in Screening shRNA Recombinant Clone

The individual bacterial colonies were adopted for templates of PCR to screen recombinant clone carrying small hairpin RNA. The PCR amplification with individual bacterial colonies and universal primer was developed to detect targeting fragment. The positive colonies were further confirmed by sequencing. Results showed that among the six clones, the positive band in the size of 324 bp were detected on agarose gel. DNA sequencing confirmed that shRNA template was cloned into plasmid vector successfully. Individual bacterial colony PCR is a simple,rapid,reliable and efficient method for screening the recombinant clones.