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Electrophoretic human leukocyte antigen HLA ‐ DQA1 DNA typing after polymerase chain reaction amplification
Author(s) -
Barros Francisco,
Carracedo Angel,
Lareu Maria Victoria,
RodriguezCalvo Maria Sol
Publication year - 1991
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150121208
Subject(s) - polymerase chain reaction , oligonucleotide , microbiology and biotechnology , electrophoresis , typing , dna , gel electrophoresis , human leukocyte antigen , dot blot , chromatography , dna extraction , biology , chemistry , gene , genetics , antigen
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 prodes 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 contol is essential. HLA ‐ DQA1 * 1 and DQA1 * 0301 can be distinguished in homoduplexes and DQA1 * 01 subtypes, DQA1 * 201 and DQA1 * 0401 in heteroduplexes (in only 5 including DNA extraction and PCR amplification). Additional variations to those recognized using commercially available dot‐blot methods can be provided since this this procedure permits the identification of single base‐pair substitutions. In addition, this method is faster and less expensive than commercial methods.