| 摘要 |
A method for the labeling of the single stranded 3' end of a target polynucleotide molecule comprises the following steps: (a) preparing an oligonucleotide in which the 3' end of the oligonucleotide is complementary to the 3' end of the target polynucleotide, and the oligonucleotide comprises a nucleotide sequence located on the 5' side of the 3' end, the nucleotide sequence being termed "the tailing sequence"; (b) incubating the oligonucleotide and the target polynucleotide under polynucleotide extending conditions; (c) adding a polymerase and labeled tri-phosphate nucleotides to the incubation mixture of step (b), wherein the labeled nucleotides are complementary to the tailing sequence of the oligonucleotide, and allowing the labeled nucleotides to extend the 3' end of the target polynucleotide; (d) optionally removing unincorporated labeled nucleotides from the mixture; and (e) optionally dissociating the oligonucleotide from the target polynucleotide. Also disclosed are a method for detecting the presence of a restriction site in a DNA molecule, and a method and kit for screening a body fluid or tissue sample for the presence of mutated DNA.
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