Barros F, Carracedo A, Lareu M V, Rodriguez-Calvo M S
Department of Legal Medicine, Faculty of Medicine, Santiago de Compostela, Spain.
Electrophoresis. 1991 Dec;12(12):1041-5. doi: 10.1002/elps.1150121208.
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-DQA101 and DQA10301 can be distinguished in homoduplexes and DQA101 subtypes, DQA10201 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.
本文描述了一种电泳方法（十二烷基硫酸钠-聚丙烯酰胺凝胶电泳），该方法无需使用等位基因特异性寡核苷酸探针和点杂交方法即可鉴定人类白细胞抗原HLA-DQA1类型和亚型。该程序可与自动化电泳系统结合用于小型凝胶。特别推荐使用PhastSystem，因为温度控制至关重要。HLA-DQA101和DQA10301可在同型双链体中区分，DQA101亚型、DQA10201和DQA1*0401可在异型双链体中区分（包括DNA提取和PCR扩增在内仅需5小时）。由于该程序允许鉴定单碱基对替换，因此可以提供比使用市售点杂交方法所识别的更多变异。此外，该方法比商业方法更快且成本更低。
