Thermal profile with alternately raised and lowered annealing temperature improves the PCR amplification using highly degenerate primers.

Abstract

The proposed here PCR thermal profile improves the specificity and efficiency of PCR using highly degenerate primers, especially in the case of larger PCR products (around 2000 bp and more). The improvement is achieved by the use of a specific annealing temperature in the beginning cycles and the alternate lowering and raising of the annealing temperature in the subsequent cycles.