PRODUCTION OF LABELED CRNA PROBES WITHOUT VECTOR CLONING - APPLICATION TO LOCALIZATION OF BASIC FIBROBLAST GROWTH-FACTOR AND TYROSINE-HYDROXYLASE MESSENGER-RNAS BY INSITU HYBRIDIZATION HISTOCHEMISTRY

We describe an approach for generating labeled, single-stranded cRNA probes, using the polymerase chain reaction with primers containing RNA polymerase promoter sequences. Transcription reactions using the amplified products and RNA polymerases yield, for the most part, full-length products. cRNA probes for basic fibroblast growth factor and tyrosine hydroxylase which have incorporated 35S-labeled nucleotides were used successfully for in situ hybridization histochemistry. This method provides a significantly faster, less work-intensive procedure for obtaining labeled single-stranded RNA useful for nucleic acid hybridization studies. © 1993 Academic Press, Inc.