title: Temperature modulation of DHPLC analysis for detection of coexisting constitutional and mosaic sequence variants in TSC2.
abstract: Somatic mosaicism is a frequent phenomenon in Mendelian disorders that exhibit a high proportion of new mutations. However, mutant alleles present at low frequency may escape detection. We have previously shown that denaturing high-performance liquid chromatography (DHPLC) at the recommended melt temperature can detect TSC1 and TSC2 mutations in tuberous sclerosis patients with low-level somatic mosaicism, even when direct sequencing cannot identify the causative lesion. Here, we report the use of temperature modulation in DHPLC analysis to facilitate the robust detection of a mosaic mutation, N1643K, in the presence of a coexisting constitutional polymorphism.
authors.affiliation: Institute of Medical Genetics, University of Wales College of Medicine, Heath Park, Cardiff CF14 4XN, UK.
authors.forename: Emmanuel S
authors.initials: ES
authors.lastname: Antonarakis
authors.forename: Julian R
authors.initials: JR
authors.lastname: Sampson
authors.forename: Jeremy P
authors.initials: JP
authors.lastname: Cheadle
mesh_terms: D000483:Alleles; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000378:metabolism; D006801:Humans; D007962:Leukocytes / Q000378:metabolism; D009154:Mutation; D011110:Polymorphism, Genetic; D011506:Proteins / Q000737:chemistry* / Q000235:genetics; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics; D013696:Temperature; D013997:Time Factors; D000077004:Tuberous Sclerosis Complex 1 Protein; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins
chemical_list: D011506:Proteins; D012097:Repressor Proteins; C000624650:TSC1 protein, human; C000624653:TSC2 protein, human; D000077004:Tuberous Sclerosis Complex 1 Protein; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins; D004247:DNA