RNA template-specific PCR: an improved method that dramatically reduces false positives in RT-PCR.
We report a novel modification of the reverse transcription PCR method, designated RNA template-specific PCR. With this approach, the 5' end of the first strand is tagged with a unique nucleotide sequence during reverse transcription that may then be exploited to amplify preferentially RNA-derived sequences. In our hands, RNA template-specific PCR retains the sensitivity of the traditional method, but greatly reduces the frequency of false positives, and virtually eliminates carryover contamination from PCR products amplified in previous experiments.
Shuldiner
A R
AR
Johns Hopkins University, Baltimore, MD.
Tanner
K
K
Moore
C A
CA
Roth
J
J
D000818:Animals; D001483:Base Sequence; D004247:DNA; D005189:False Positive Reactions; D007328:Insulin / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000032:analysis*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D013698:Templates, Genetic; D014981:Xenopus
D007328:Insulin; D012313:RNA; D004247:DNA; D012194:RNA-Directed DNA Polymerase