UNIT 5.13 Fluorescent In Situ Transcription in Cells and Tissues

  1. Marjorie A. Ariano

Published Online: 1 MAY 2001

DOI: 10.1002/0471142301.ns0513s08

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Ariano, M. A. 2001. Fluorescent In Situ Transcription in Cells and Tissues. Current Protocols in Neuroscience. 8:5.13:5.13.1–5.13.10.

Author Information

  1. The Chicago Medical School, North Chicago, Illinois

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: AUG 1999


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.