1. Complementary strand analysis: a new approach for allelic separation in complex polyallelic genetic systems.
- Author
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Arguello R, Pay AL, McDermott A, Ross J, Dunn P, Avakian H, Little AM, Goldman J, and Madrigal JA
- Subjects
- Electrophoresis, Polyacrylamide Gel, Female, Genetic Complementation Test, HLA-A Antigens genetics, HLA-B Antigens genetics, HLA-C Antigens genetics, Humans, Male, Polymerase Chain Reaction, Polymorphism, Genetic, Alleles, Genetic Techniques
- Abstract
We describe a method, complementary strand analysis (CSA), for separating alleles potentially from any heterozygous genetic locus. Locus specific PCR is performed generating two allelic products. The antisense strands are isolated and hybridised with a sense reference strand to form a chimeric DNA duplex for each allele which is then separated by non-denaturing PAGE. We demonstrate the application of CSA for separation of highly polymorphic HLA-A, -B and -Cw alleles and characterisation of HLA identity in related bone marrow donors and patients. CSA is capable of resolving one nucleotide differences in a DNA fragment nearly as large as a kilobase in length.
- Published
- 1997
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