No 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.

The 3' part of the glucosamine-6-phosphate synthase gene from Histoplasma capsulatum was PCR amplified using degenerate primers designed from the known glucosamine-6-phosphate synthase gene sequences, cloned and sequenced. The computer analysis of the 676 bp sequence revealed the presence of two introns. The identities of the deduced amino acid sequence to the corresponding Saccharomyces cerevisiae and Candida albicans fragment are 65 and 63.8%, respectively.