Dideoxy DNA sequencing with end-labeled oligonucleotide primers.
End-labeled oligonucleotide primers may be used effectively as the source of radiolabel in DNA sequencing by the dideoxy method. The approach is demonstrated with various end-labeled oligonucleotides, including a commercially prepared universal primer and a mixed-sequence probe. Single-stranded (M13) or denatured double-stranded template may be used. The end-labeled primers and their extension products may be stored for weeks. The method is useful for identification of clones isolated by oligonucleotide hybridization and it provides a convenient, economical alternative to the use of alpha-labeled deoxynucleoside triphosphate.
D001483:Base Sequence; D004247:DNA; D004277:DNA, Single-Stranded; D004229:Dithiothreitol; D009691:Nucleic Acid Denaturation; D009841:Oligonucleotides; D013698:Templates, Genetic
D004277:DNA, Single-Stranded; D009841:Oligonucleotides; D004247:DNA; D004229:Dithiothreitol