Polyvinyl alcohol enhances detection of low abundance transcripts in early stage quail embryos in a nonradioactive whole mount in situ hybridization technique

Biotechniques. 1994 Aug;17(2):324, 326-7.

Abstract

We have tested the ability of polyvinyl alcohol (PVA) to enhance the indoxyl-nitro blue tetrazolium alkaline phosphatase color reaction in whole mount in situ hybridizations of early stage quail embryos. Using detection of the myogenic factor qmyoD in hybridizations of 13-14 somite-pair quail embryos as a test system, 10% solutions of 13-23 kDa and 31-50 kDa PVA optimally enhanced the alkaline phosphatase color reaction. With the enhanced detection conditions, we have been able to measure qmyoD expression as early as the 3 somite-pair stage, whereas previously we were unable to detect qmyoD prior to the 10 somite-pair stage. PVA-enhanced detection of somitic qmyoD expression was specific as no corresponding signal was found in control hybridizations with a sense qmyoD probe.

Publication types

  • Research Support, Non-U.S. Gov't
  • Technical Report

MeSH terms

  • Animals
  • Coturnix / embryology*
  • Female
  • In Situ Hybridization / methods*
  • Myogenic Regulatory Factors / genetics*
  • Polyvinyl Alcohol / pharmacology*
  • RNA, Messenger / analysis*

Substances

  • Myogenic Regulatory Factors
  • RNA, Messenger
  • Polyvinyl Alcohol