Electrophoretic human leukocyte antigen HLA-DQA1 DNA typing after polymerase chain reaction amplification

Electrophoresis. 1991 Dec;12(12):1041-5. doi: 10.1002/elps.1150121208.

Abstract

An electrophoretic method (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) is described which permits the identification of human leukocyte antigen HLA-DQA1 types and subtypes without using allele-specific oligonucleotide probes and dot-blot methodology. The procedure can be used in miniaturized gels in combination with automated electrophoretic systems. The PhastSystem is particularly recommended since temperature control is essential. HLA-DQA1*01 and DQA1*0301 can be distinguished in homoduplexes and DQA1*01 subtypes, DQA1*0201 and DQA1*0401 in heteroduplexes (in only 5 h including DNA extraction and PCR amplification). Additional variations to those recognized using commercially available dot-blot methods can be provided since this procedure permits the identification of single base-pair substitutions. In addition, this method is faster and less expensive than commercial methods.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA / genetics*
  • Electrophoresis, Polyacrylamide Gel / methods*
  • HLA-DQ Antigens / genetics*
  • HLA-DQ alpha-Chains
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction*
  • Silver Staining

Substances

  • HLA-DQ Antigens
  • HLA-DQ alpha-Chains
  • HLA-DQA1 antigen
  • DNA