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.
Figeys
D
D
Department of Chemistry, University of Alberta, Edmonton, Canada.
Renborg
A
A
Dovichi
N J
NJ
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