RNA template-specific PCR: an improved method that dramatically reduces false positives in RT-PCR

Biotechniques. 1991 Dec;11(6):760-3.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA
  • False Positive Reactions
  • Insulin / genetics
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods*
  • RNA / analysis*
  • RNA-Directed DNA Polymerase / metabolism
  • Templates, Genetic*
  • Xenopus

Substances

  • Insulin
  • RNA
  • DNA
  • RNA-Directed DNA Polymerase