title: Complementary strand analysis: a new approach for allelic separation in complex polyallelic genetic systems.
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.
authors.affiliation: The Anthony Nolan Research Institute, The Royal Free Hospital, Pond Street, Hampstead, London NW3 2QG, UK.
authors.forename: R
authors.initials: R
authors.lastname: Arguello
authors.forename: A L
authors.initials: AL
authors.lastname: Pay
authors.forename: A
authors.initials: A
authors.lastname: McDermott
authors.forename: J
authors.initials: J
authors.lastname: Ross
authors.forename: P
authors.initials: P
authors.lastname: Dunn
authors.forename: H
authors.initials: H
authors.lastname: Avakian
authors.forename: A M
authors.initials: AM
authors.lastname: Little
authors.forename: J
authors.initials: J
authors.lastname: Goldman
authors.forename: J A
authors.initials: JA
authors.lastname: Madrigal
mesh_terms: D000483:Alleles; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005816:Genetic Complementation Test; D005821:Genetic Techniques; D015234:HLA-A Antigens / Q000235:genetics; D015235:HLA-B Antigens / Q000235:genetics; D015236:HLA-C Antigens / Q000235:genetics; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic
chemical_list: D015234:HLA-A Antigens; D015235:HLA-B Antigens; D015236:HLA-C Antigens