A reliable method for northern blot analysis using synthetic oligonucleotide probes

Biotechniques. 1991 Feb;10(2):190-7.

Abstract

We have developed a method for using short (30-42 base pair) synthetic oligonucleotide DNA probes in Northern blot assays. The method involves labeling the probes to high specific activity, very stringent hybridization and wash conditions, and the presence of several inhibitors of nonspecific binding in the hybridization buffer. We have tested this method with several probes obtained from local and commercial sources. The results with every probe used were high signal-to-noise ratios in an exposure time range of 30 min to 7 days.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Northern / methods*
  • Cell Line
  • DNA Probes*
  • Humans
  • Lymphokines / genetics
  • Mice
  • Mice, Inbred CBA
  • Oligonucleotide Probes*
  • Prealbumin / genetics
  • RNA / analysis
  • Schistosomiasis mansoni / genetics

Substances

  • DNA Probes
  • Lymphokines
  • Oligonucleotide Probes
  • Prealbumin
  • RNA