Quantification of specific mRNA by flatbed scintillation counting of dual-labeled dot blots

Biotechniques. 1993 Oct;15(4):738-43.


A dual-labeling technique was developed for direct quantification of specific mRNA using a flatbed liquid scintillation counter. This method simultaneously measures cpm of 32P- and 35S-labeled probes bound to RNA dot blots and subtracts counts due to nonspecific background radioactivity bound to the filter. Probes for T-cell receptor and beta-actin (as the internal standard) were hybridized both separately and simultaneously to RNA isolated from five different sources. There was concordance between the radioactivity measured from single- and dual-hybridizations for each combination of 35S- and 32P-labeled probes. This methodology directly quantifies specific mRNA sequences bound to membranes and has potential for measuring gene dosage, without the need for re-probing or densitometric analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics
  • Animals
  • Filtration / instrumentation
  • Mice
  • Nucleic Acid Hybridization
  • Phosphorus Radioisotopes
  • RNA Probes
  • RNA, Messenger / analysis*
  • Receptors, Antigen, T-Cell / genetics
  • Scintillation Counting*
  • Sulfur Radioisotopes


  • Actins
  • Phosphorus Radioisotopes
  • RNA Probes
  • RNA, Messenger
  • Receptors, Antigen, T-Cell
  • Sulfur Radioisotopes