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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
Author(s) -
Rolfs Becky K.,
Wu Ching Ching,
Lerche Nicholas W.,
Smith David G.
Publication year - 2001
Publication title -
american journal of physical anthropology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.146
H-Index - 119
eISSN - 1096-8644
pISSN - 0002-9483
DOI - 10.1002/ajpa.1126
Subject(s) - genotyping , typing , biology , genotype , genetics , allele , gene
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. Am J Phys Anthropol 116:296–301, 2001. © 2001 Wiley‐Liss, Inc.