Complementary strand analysis: a new approach for allelic separation in complex polyallelic genetic systems.
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.
Arguello
R
R
The Anthony Nolan Research Institute, The Royal Free Hospital, Pond Street, Hampstead, London NW3 2QG, UK.
Pay
A L
AL
McDermott
A
A
Ross
J
J
Dunn
P
P
Avakian
H
H
Little
A M
AM
Goldman
J
J
Madrigal
J A
JA
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
D015234:HLA-A Antigens; D015235:HLA-B Antigens; D015236:HLA-C Antigens