title: Quantitative real-time PCR eliminates false-positives in colony screening PCR.
abstract: We report an alternative approach to colony screening using real-time PCR (qPCR) which can be used instead of the traditional end-point PCR to eliminate false-positives and reduce processing times. False-positive transformants can easily be distinguished from true-positives by comparing Ct values derived from qPCR amplification curves. In addition, the use of qPCR allows for more efficient processing since a gel electrophoresis step is not required and the screening process is no longer limited by the capacity of the gel apparatus.
authors.affiliation: Sydney Medical School Nepean, University of Sydney, Nepean Hospital, Penrith, New South Wales, Australia.
authors.forename: Kristen K
authors.initials: KK
authors.lastname: Skarratt
authors.affiliation: Sydney Medical School Nepean, University of Sydney, Nepean Hospital, Penrith, New South Wales, Australia. Electronic address: stephen.fuller@sydney.edu.au.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Fuller
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D005189:False Positive Reactions; D005820:Genetic Testing / Q000379:methods; D005827:Genetics, Microbial / Q000379:methods; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D014169:Transformation, Bacterial
keywords: Colony PCR; False-positive; Transformant; qPCR