title: Reverse transcription and direct amplification of cellular RNA transcripts by Taq polymerase.
abstract: We report the ability of Taq polymerase to directly transcribe RNA templates in vitro. We have made use of this finding to develop a single-step protocol for amplification of RNA transcripts. The method was shown to require only subnanogram amounts of total cellular RNA as starting material. A microassay was developed in which RNA can be extracted from one drop of blood or 1000 cultured cells, and analyzed for the expression of a specific gene.
authors.affiliation: Department of Human Genetics, Yale University School of Medicine, New Haven, CT 06510.
authors.forename: W T
authors.initials: WT
authors.lastname: Tse
authors.forename: B G
authors.initials: BG
authors.lastname: Forget
mesh_terms: D001483:Base Sequence; D004274:DNA, Recombinant; D004259:DNA-Directed DNA Polymerase; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger; D013049:Spectrin / Q000235:genetics; D019914:Taq Polymerase; D013698:Templates, Genetic; D014158:Transcription, Genetic
chemical_list: D004274:DNA, Recombinant; D012333:RNA, Messenger; D013049:Spectrin; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase