title: RNA Template-Specific Polymerase Chain Reaction (RS-PCR) : A Modification of RNA-PCR that Dramatically Reduces the Frequency of False Positives.
abstract: Reverse transcription of RNA followed by the polymerase chain reaction (RT-PCR or RNA-PCR) is an extraordinarily sensitive method to detect as few as 1-100 copies of a specific RNA (1-3). However, we and others have found that false positives caused by contamination with minute quantities of DNA (i.e., cDNAs, plasmid DNAs, genomic DNA, or PCR carryover) is a major shortcoming of the method even when meticulous laboratory technique is employed (4-6). RNA template-specific PCR (RS-PCR) is a modification of conventional RNA-PCR in which RNA is reverse-transcribed with a primer that contains at its 5' end a unique nucleotide sequence that may then be exploited in the PCR to amplify preferentially the RNA-derived sequence. RS-PCR retains full sensitivity, but reduces dramatically the frequency of false positives (7-9).
authors.affiliation: Department of Medicine, Johns Hopkins University School of Medicine and National Institute on Aging, National Institutes of Health, Baltimore, MD.
authors.forename: A R
authors.initials: AR
authors.lastname: Shuldiner
authors.forename: R
authors.initials: R
authors.lastname: Perfetti
authors.forename: J
authors.initials: J
authors.lastname: Roth