Fluorescent in situ transcription in cells and tissues.
This method assesses cellular mRNA transcripts in tissue sections and cell cultures using unique short anti-sense primers directed against sequences in particular protein(s). The unlabeled synthetic cDNA oligonucleotide primers are extended complementary to a sense mRNA transcript using reverse transcriptase and labeled through incorporation of a fluorescent-labeled dUTP nucleotide base. The new cDNA will be synthesized upstream from the point of primer hybridization, and has a specific activity of fluorescent labeling dependent upon the length of the template mRNA from the primer location to the 5'-terminus. This procedure provides rapid detection of low abundance mRNA messages that can be related to other cellular protein components, labeled experimentally with alternative fluorochromes.
Ariano
M A
MA
The Chicago Medical School, North Chicago, Illinois, USA.
D000818:Animals; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics*
D012333:RNA, Messenger