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 adequate 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 enhance the ability to successfully genotype DNA from samples that were of poor quality and/or quantity. Long-term storage of whole-blood buffy coats or stored DNA extracted from whole-blood buffy coats did not affect typing success; however, amplification and typing of DNA extracted from archival samples of plasma were difficult and resulted in a low success rate. This suggests that amplification and DQA1-genotyping of archival samples is possible with a modified protocol, but is influenced by the age and source of the sample, and to a lesser extent, the method used to extract DNA from sample substrates.
Rolfs
B K
BK
Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California at Davis, Davis, California 95616, USA. bkrolfs@msn.com
Wu
C C
CC
Lerche
N W
NW
Smith
D G
DG
D000818:Animals; D005784:Gene Amplification; D005802:Genes, MHC Class II / Q000235:genetics*; D008253:Macaca mulatta / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D013048:Specimen Handling; D013997:Time Factors