Suppression of pyrophosphorolysis in DNA sequencing and in other

Details Suppression of pyrophosphorolysis in DNA sequencing and in other Suppression of pyrophosphorolysis in DNA sequencing and in other

1997-07-23

1999-02-16

Arthur, Lisa B.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

436 94, 935 77, 935 78, C12Q 168, G01N 3348

Patent

active

058719296

ABSTRACT:
A method of inhibiting pyrophosphorolysis during DNA chain length elongation is provided. The method comprises including both Mn.sup.++ and Mg.sup.++ in chain-extension and chain-termination reaction mixtures such as those used in DNA sequencing. This method is useful in stabilizing dideoxy-ribonucleoside triphosphate-terminated DNA chains and improving the quality of DNA sequence data obtained via the use of DNA polymerases that do not discriminate against the incorporation of dideoxyribonucleoside triphosphates and other chain terminating agents. Also provided are a reaction mixture and kit for DNA sequencing employing this method.

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