title: Production of Labeled cRNA Probes without Vector Cloning: Application to Localization of Basic Fibroblast Growth Factor and Tyrosine Hydroxylase mRNAs by in Situ Hybridization Histochemistry.
abstract: 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 (35) S-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.
authors.affiliation: Department of Histology and Neurobiology, Karolinska Institute, Stockholm, S-104 01, Sweden; and Ludwig Institute for Cancer Research, Stockholm, Sweden.
authors.forename: A J
authors.initials: AJ
authors.lastname: Bean
authors.forename: J F
authors.initials: JF
authors.lastname: Simons
authors.forename: T
authors.initials: T
authors.lastname: Hökfelt