Abstract
Quantitative real-time PCR (qPCR) was applied to rapid screening of positive plasmid clones. Insert-specific primer pairs were used in qPCR colony screening, and false positive colonies could easily be distinguished from true positive ones by comparing their Ct values. In addition, qPCR is particularly suitable when amplicon is small (<150 bp). This method is sensitive, simple and fast, obviates the need for gel electrophoresis, and is a cost-effective alternative to the traditional PCR approach.
Highlights
Open Access sential for many experimental purposes such as DNA amplification, DNA library construction, protein expression, ShRNA gene knockdown, and in the recent CRISPR/Cas
The resulting transformed E. coli grown on appropriate selective medium contains recombinant plasmids or self-ligated vector colonies, which are subsequently screened for colonies containing the desired DNA insert with the right orientation [2]
Plasmid cloning is characterized by picking bacterial colonies, growing bacteria in selective medium overnight, preparing plasmid DNA, and digesting with restriction enzymes
Summary
Open Access sential for many experimental purposes such as DNA amplification, DNA library construction, protein expression, ShRNA gene knockdown, and in the recent CRISPR/Cas. Quantitative real-time PCR (qPCR) was applied to rapid screening of positive plas- Screening of Recombinant Plasmids by Direct Colony Quantitative Real-Time PCR.
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call