Major histocompatibility complex class II polymorphisms in Macaca mulatta: Factors influencing comprehensive genotyping of Macaca mulatta (Mamu)- DQA1 alleles by PCR-RFLP in archival samples

In the last 5 years, HLA class II genotyping methods have been adapted for genotyping of class II loci in rhesus macaques. Since previously published typing protocols were used on samples that were collected and stored under ideal conditions, it was of interest to determine if these methods were ade quate for genotyping a large collection of archival samples from which DNA had been isolated and stored under various conditions. Established macaque DQA1 typing protocols were modified to optimize the typing procedure and en hance the ability to successfully genotype DNA from samples that were of po or quality and/or quantity. Long-term storage of whole-blood huffy coats or stored DNA extracted from whole-blood buffy coats did not affect typing su ccess; however, amplification and typing of DNA extracted from archival sam ples of plasma were difficult and resulted in a low success rate. This sugg ests that amplification and DQA1-genotyping of archival samples is possible with a modified protocol, but is influenced by the age and source of the s ample, and to a lesser extent, the method used to extract DNA from sample s ubstrates. (C) 2001 Wiley-Liss, Inc.