title: Synthesis and automated detection of fluorescently labeled primer extension products.
abstract: Common methods for determination of transcription initiation sites are based on the extension of radioactively labeled primers or internal labeling of extension products with reverse transcriptase. Here, we describe a modification of this procedure, adapted for automated detection of extension products by using fluorescent primers and an optimized reaction protocol. This facilitates high-throughput screening to detect mRNA start-points of large DNA regions and significantly reduces the time required to determine sites of transcription initiation.
authors.affiliation: Abt. Mikrobiologie, Universität Kaiserslautern, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Altermann
authors.forename: J R
authors.initials: JR
authors.lastname: Klein
authors.forename: B
authors.initials: B
authors.lastname: Henrich
mesh_terms: D001435:Bacteriophages / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers; D005456:Fluorescent Dyes; D007778:Lactobacillus / Q000821:virology; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics
chemical_list: D017931:DNA Primers; D005456:Fluorescent Dyes; D012329:RNA, Bacterial; D012367:RNA, Viral; D014764:Viral Proteins; C111930:gporf2 protein, bacteriophage phiadh