Polyvinyl alcohol enhances detection of low abundance transcripts in early stage quail embryos in a nonradioactive whole mount in situ hybridization technique.
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.
Barth
J
J
University of Georgia, Athens 30602.
Ivarie
R
R
D000818:Animals; D003370:Coturnix / Q000196:embryology*; D005260:Female; D017403:In Situ Hybridization / Q000379:methods*; D018006:Myogenic Regulatory Factors / Q000235:genetics*; D011142:Polyvinyl Alcohol / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis*
D018006:Myogenic Regulatory Factors; D012333:RNA, Messenger; D011142:Polyvinyl Alcohol