Mutation detection and typing of polymorphic loci through double-strand conformation analysis
- 1 February 1998
- journal article
- Published by Springer Science and Business Media LLC in Nature Genetics
- Vol. 18 (2), 192-194
- https://doi.org/10.1038/ng0298-192
Abstract
Variations, such as nucleotide substitutions, deletions and insertions, within genes can affect the function of the gene product and in some cases be deleterious. Screening for known allelic variation is important for determining disease and gene associations. Techniques which target specific mutations such as restriction enzyme polymorphism and oligonucleotide probe or PCR primer reactivity are useful for the detection of specific mutations, but these techniques are not generally effective for the identification of new mutations. Approaches for measuring changes in DNA conformation have been developed, based on the principle that DNA fragments which differ in nucleotide composition exhibit different mobilities after separation by polyacrylamide gel electrophoresis (PAGE). Here we describe a conformation-based mutation detection system, double-strand conformation analysis (DSCA), which provides a simple means to detect genetic variants and to type complex polymorphic loci. We demonstrate the application of DSCA to detect genetic polymorphisms such as a single-nucleotide difference within DNA fragments of up to 979 base pairs in length. We present the application of DSCA in detecting four different mutations in the cystic fibrosis gene (CFTR) and 131 different alleles encoded by HLA class I genes.Keywords
This publication has 10 references indexed in Scilit:
- Complementary strand analysis: a new approach for allelic separation in complex polyallelic genetic systemsNucleic Acids Research, 1997
- Genes, disease and medicineBritish Journal of Clinical Pharmacology, 1996
- A rapid HLA‐DRB1*04 subtyping method using PCR and DNA heteroduplex generatorsTissue Antigens, 1996
- Complete subtyping of the HLA‐A locus by sequence‐specific amplification followed by direct sequencing or single‐strand conformation polymorphism analysisTissue Antigens, 1995
- Locus‐specific amplification of HLA class I genes from genomic DNA: locus‐specific sequences in the first and third introns of HLA‐A, ‐B, and ‐C allelesTissue Antigens, 1995
- Retrospective study of the cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations in Guthrie cards from a large cohort of neonatal screening for cystic fibrosisHuman Genetics, 1994
- Exploiting structural differences among heteroduplex molecules to simplify genotyping the DQA1 and DQB1 alleles in human lymphocyte typingNucleic Acids Research, 1993
- Genomic DNA sequence of the cystic fibrosis transmembrane conductance regulator (CFTR) geneGenomics, 1991
- Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms.Proceedings of the National Academy of Sciences, 1989
- Detection of single base substitutions in total genomic DNANature, 1985