Patent ID: 7150980
Filing Date: 2006-12-19
Classification: C12Q

Abstract:
1. In a method for performing a cycled primer extension reaction using a non-thermostable polymerase, comprising: (i) contacting a template DNA comprising a target sequence of nucleotides with at least one primer oligonucleotide complementary to a nucleotide sequence at the 3′-end of said target sequence, under conditions allowing annealing of said primer to its complementary nucleotide sequence on said target sequence; and (ii) carrying out a polymerase-mediated extension of said primer on said target sequence of nucleotides, the improvement wherein the non-thermostable polymerase is stabilized and the operating temperatures are optimized for non-thermostable polymerase at temperatures where the non-thermostable polymerase would otherwise be inactivated, comprising: conducting the cycled primer extension reaction in the presence of a sufficient concentration of proline to allow operating temperatures at which the non-thermostable polymerase is stabilized and optimized.