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Mutation analysis of the hamartin gene using denaturing high performance liquid chromatography
Author(s) -
Bénit Paule,
KaraMostefa Ali,
Berthelon Monique,
Sengmany Kamchan,
Munnich Arnold,
Bonnefont JeanPaul
Publication year - 2000
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/1098-1004(200011)16:5<417::aid-humu6>3.0.co;2-4
Subject(s) - biology , denaturing high performance liquid chromatography , mutation , gene , genetics , microbiology and biotechnology
Denaturing high performance liquid chromatography (DHPLC) is a novel high‐capacity technique for gene mutation scanning. We have assessed the sensitivity and specificity of this method for analysis of the full coding sequence of the hamartin (TSC1) gene in 20 tuberous sclerosis patients, whose TSC1 genes previously had been studied by single strand conformation polymorphism analysis and protein truncation assay. All eight sequence variants previously identified were adequately detected by DHPLC. Additionally, this approach picked up three polymorphisms, one of which (IVS13‐55 C>G) was hitherto unreported, therefore serving as proof of principle for this technique. Thus, DHPLC appears to be a highly sensitive method with advantages in terms of flexibility, fragments size analysis, cost and time and labor sparing, compared to classical approaches of mutation scanning. Hum Mutat 16:417–421, 2000. © 2000 Wiley‐Liss, Inc.