Labeling of double-stranded DNA by ROX-dideoxycytosine triphosphate using terminal deoxynucleotidyl transferase and separation by capillary electrophoresis.
Terminal transferase is used to add a single fluorescently labeled dideoxynucleotide to double-stranded DNA prepared by restriction endonuclease action on a bacteriophage. The product is separated by capillary electrophoresis with both hydroxypropylmethylcellulose and non-cross-linked polyacrylamide. The reaction products generate single peaks for each fragment with hydroxypropylmethylcellulose. However, the higher resolution separation produced by non-cross-linked polyacrylamide shows that the product contains two components for each restriction digest fragment. This labeling technique should be useful in restriction fragment length polymorphism studies.
Department of Chemistry, University of Alberta, Edmonton, Canada.
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D
Figeys
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A
Renborg
N J
NJ
Dovichi
D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004253:DNA Nucleotidylexotransferase / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D003845:Deoxycytosine Nucleotides / Q000737:chemistry*; D054306:Dideoxynucleotides; D004586:Electrophoresis; D005456:Fluorescent Dyes; D065347:Hypromellose Derivatives; D008747:Methylcellulose / Q000031:analogs & derivatives / Q000737:chemistry; D011110:Polymorphism, Genetic; D013050:Spectrometry, Fluorescence
D004268:DNA-Binding Proteins; D003845:Deoxycytosine Nucleotides; D054306:Dideoxynucleotides; D005456:Fluorescent Dyes; D065347:Hypromellose Derivatives; C037406:2',3'-dideoxycytidine 5'-triphosphate; D008747:Methylcellulose; D004247:DNA; D004253:DNA Nucleotidylexotransferase