Asymmetric polymerase chain reaction-single-strand conformation polymorphism (asymmetric PCR-SSCP) as a simple method for allele typing of HLA-DRB

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Title
Asymmetric polymerase chain reaction-single-strand conformation polymorphism (asymmetric PCR-SSCP) as a simple method for allele typing of HLA-DRB
Author(s)
Joo Hyun Kang; Kyeong Hee Kim; Cheol Young Maeng; Kil Lyong Kim
Bibliographic Citation
BMB Reports, vol. 32, no. 6, pp. 529-534
Publication Year
1999
Abstract
Asymmetric PCR and single-strand conformation polymorphism (SSCP) methods were combined to analyze human leukocyte antigen (HLA)-DRB allele polymorphism. Asymmetric PCR amplification was applied to generate single-stranded DNA (ssDNA) using the nonradioactive oligonucleotide primers designed for the polymorphic exon 2 region. The conformational differences of ssDNAs, depending on the allele type, were analyzed by nondenaturing polyacrylamide gel electrophoresis and visualized by ethidium bromide staining. The ssDNAs were clearly separated from double-stranded DNA without interference and obviously migrated depending on their allele type. This method was applied to the genomic DNA either from homozygous or from heterozygous cell lines containing the DR4 allele as template DNA using DR4-specific primers, and satisfying results were obtained. Compared to the standard PCR-SSCP method, this asymmetric PCR-SSCP method has advantages of increased speed, reproducibility, and convenience. Along with PCR-SSP or sequence-based typing, this method will be useful in routine typing of HLA-DRB allele.
Keyword
Asymmetric PCRHLA class II DR moleculePolymorphismSingle-Strand Conformation Polymorphism
ISSN
1225-8687
Publisher
Korea Soc-Assoc-Inst
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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