A method for typing polymorphism at the HLA‐A locus using PCR amplification and immobilized oligonucleotide probes
作者： Teodorica L. Bugawan , Raymond Apple , Henry A. Erlich
DOI: 10.1111/J.1399-0039.1994.TB02371.X
关键词:
摘要: We have developed a simple and rapid method for DNA typing of the HLA-A locus using PCR amplification hybridization product, labeled with biotinylated primers, to an array immobilized oligonucleotide probes in single reaction (reverse dot or line blot). A primer set (RAP1007 DB337) is used specifically amplify 990-bp fragment containing exons 1, 2, 3 from genomic DNA. This locus-specific amplifies all alleles. 51 sequence-specific (SSO) probes, 25 exon 2 26 3, was nylon membrane by UV-crosslinking poly-thymidine "tail" added terminal transferase. In blot format, 50 SSO plus control probe are on strip. The amplified variety samples. These can identify 37 homozygous analysis heterozygous samples, 604 types out 633 (95.4%) possible patterns be detected as unique reactivity pattern. computer program has been assign alleles genotypes based
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