Method and kit for polynucleotide assay including primer-dependant DNA polymerase
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United States of America Patent
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Jul 25, 1989
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N/A
app pub date -
May 16, 1986
filing date -
May 16, 1986
priority date (Note) -
Expired
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Abstract
A probe polynucleotide binds to a target nucleotide sequence in the nucleic acid of a biological sample, and then is enzymatically extended in the 3'-direction with a mixture of nucleoside triphosphates including at least one nucleoside triphosphate that has been detectably labeled. After separating extended hybrid from unreacted nucleoside triphosphates, detectably-modified nucleotides which have been incorporated are determined. In some forms, the 3'-terminal nucleotide of the probe polynucleotide is selected to form a matched pair with some sample strands, but a mismatched pair with other sample strands. In such cases, if the primer dependent enzyme used for extension is one lacking 3'-exonuclease activity, then only those hybrids forming such a matched pair will be extended and subsequently determined.
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