Construction of recombinant RNA templates for use as internal standards in quantitative RT-PCR

John Patrick Vanden Heuvel, F. L. Tyson, D. A. Bell

Research output: Contribution to journalArticle

179 Scopus citations

Abstract

The PCR has proven to be useful in the analysis of gene expression of specific mRNAs. Although PCR is able to detect rare mRNA transcripts following reverse transcription PCR, determining relative or absolute copy number can be difficult due to sample-to-sample variation. The use of a recombinant mRNA internal standard that contains target mRNA primer sequences greatly improves reproducibility of quantitation. Reverse transcription PCR products generated from the internal standard can be distinguished from the product generated from the target gene mRNA because of their size differences. In this report we present a facile and general PCR-based method for synthesis of internal standards that may be used as competitive or co-amplified templates for quantitative reverse transcription PCR.

Original languageEnglish (US)
Pages (from-to)395-398
Number of pages4
JournalBioTechniques
Volume14
Issue number3
StatePublished - Jan 1 1993

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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