Synthesis and automated detection of fluorescently labeled primer extension products.
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.
Altermann
E
E
Abt. Mikrobiologie, Universität Kaiserslautern, Germany.
Klein
J R
JR
Henrich
B
B
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
D017931:DNA Primers; D005456:Fluorescent Dyes; D012329:RNA, Bacterial; D012367:RNA, Viral; D014764:Viral Proteins; C111930:gporf2 protein, bacteriophage phiadh