Synthesis of digoxigenin-labeled cDNA as a test for mRNA integrity

Biotechniques. 1992 Dec;13(6):876-9.

Abstract

The nucleotide analogue digoxigenin-11-dUTP is widely used as a nonradioactive marker in a broad range of techniques including dot blots, Southern and Northern blots, colony and plaque screenings and in situ hybridizations. In this report, we describe the incorporation of this molecule into a cDNA synthesized by reverse transcriptase as a novel application for digoxigenin. This reaction can be performed as a useful control to check the integrity of a bulk mRNA preparation in the construction of a cDNA library, since the ability of mRNA to direct the synthesis of long molecules of first-strand cDNA is a sign of its integrity.

MeSH terms

  • DNA / biosynthesis*
  • DNA / genetics
  • DNA Probes
  • Deoxyuracil Nucleotides*
  • Digoxigenin / analogs & derivatives*
  • Evaluation Studies as Topic
  • Genetic Techniques
  • Molecular Probe Techniques
  • RNA, Messenger / genetics*
  • RNA-Directed DNA Polymerase / metabolism

Substances

  • DNA Probes
  • Deoxyuracil Nucleotides
  • RNA, Messenger
  • digoxigenin-11-deoxyuridine triphosphate
  • DNA
  • RNA-Directed DNA Polymerase
  • Digoxigenin