Suppression of pyrophosphorolysis in DNA sequencing and in other applications involving DNA replication

摘要

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.