Comparative method for detection of RNA-PCR-amplified signals

Biotechniques. 1992 Jan;12(1):22, 24, 26.

Abstract

During the development of a micro-method for the quantitative analysis of gene expression, we observed that the sensitivity for detection of PCR-amplified product was higher when silver staining was used, compared with either ethidium bromide staining (as reported previously) or Southern hybridization with 32P-labeled oligonucleotide probe. This observation may have an important impact on the analysis of RNA-PCR-amplified signal for the quantitation of mRNA expression, simply by densitometric scanning of silver-stained polyacrylamide gel. We believe that this is the first report to show such a comparison, demonstrating the greater sensitivity of silver staining compared with either ethidium bromide staining or Southern hybridization utilizing an oligonucleotide probe prepared by 3'-end labeling with 32P-dATP.

Publication types

  • Comparative Study

MeSH terms

  • Base Sequence
  • Blotting, Southern*
  • Breast Neoplasms / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Ethidium
  • Gene Expression*
  • Humans
  • Molecular Sequence Data
  • Oligonucleotide Probes
  • Polymerase Chain Reaction*
  • RNA, Messenger / genetics
  • RNA, Neoplasm / analysis*
  • RNA, Neoplasm / genetics
  • Silver*
  • Staining and Labeling*

Substances

  • Oligonucleotide Probes
  • RNA, Messenger
  • RNA, Neoplasm
  • Silver
  • Ethidium