Patent ID: 6428986
Filing Date: 2002-08-06
Classification: C12Q

Abstract:
A method for performing a cycled primer extension reaction comprising the steps of:(i) contacting a template DNA comprising a target sequence of nucleotides with at least one primer oligonucleotide complementary to a nucleotide sequence at the 3â€²-end of said target sequence, under conditions allowing annealing of said prior to its complementary nucleotide sequence on said target sequence, in the presence of an osmoprotectant selected from 2-methyl-4-carboxy-3,4,5,6-tetrahydropyrimidine and mixtures of THP(B) and proline, to lower the melting temperature of said template DNA and/or of said primer; and (ii) carrying out a polymerase-mediated extension of said primer on said target sequence of nucleotides in the presence of an osmoprotectant selected from THP(B), 2-methyl-4-carboxy-5-hydroxy-3,4,5,6-tetrahydropyrimidine and mixtures of THP(B), THP(A), and/or proline to stabilize said polymerase; thus obtaining a high yield specific extension of the primer on said target sequence of nucleotides of the template DNA.