Synthesis of RNA probes by the direct in vitro transcription of PCR-generated DNA templates

Raul Urrutia, Mark A. McNiven, Bechara Kachar

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

We describe a novel method for the generation of RNA probes based on the direct in vitro transcription of DNA templates amplified by polymerase chain reaction (PCR) using primers with sequence hybrids between the target gene and those of the T7 and T3 RNA polymerases promoters. This method circumvents the need for cloning and allows rapid generation of strand-specific RNA molecules that can be used for the identification of genes in hybridization experiments. We have successfully applied this method to the identification of DNA sequences by Southern blot analysis and library screening.

Original languageEnglish (US)
Pages (from-to)113-120
Number of pages8
JournalJournal of Biochemical and Biophysical Methods
Volume26
Issue number2-3
DOIs
StatePublished - May 1993

Keywords

  • Hybridization
  • In vitro transcription
  • Kinesin
  • Polymerase chain reaction
  • RNA probe

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

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