title: Allele-Specific DNA Methylation Detection by Pyrosequencing®.
abstract: DNA methylation is an epigenetic modification that plays important roles in healthy as well as diseased cells, by influencing the transcription of genes. In spite the fact that human somatic cells are diploid, most of the currently available methods for the study of DNA methylation do not provide information on the methylation status of individual alleles of genes. This information may be of importance in many situations. In particular, in cancer both alleles of tumour suppressor genes generally need to be inactivated for a phenotypic effect to be observed. Here, we present a simple and cost-effective protocol for allele-specific DNA methylation detection based on Pyrosequencing(®) of methylation-specific PCR (MSP) products including a single nucleotide polymorphism (SNP) within the amplicon.
authors.affiliation: Department of Hematology, Rigshospitalet, Blegdamsvej 9, 2100-DK, Copenhagen, Denmark, lasse.sommer.kristensen@regionh.dk.
authors.forename: Lasse Sommer
authors.initials: LS
authors.lastname: Kristensen
authors.forename: Jens Vilstrup
authors.initials: JV
authors.lastname: Johansen
authors.forename: Kirsten
authors.initials: K
authors.lastname: Grønbæk
mesh_terms: D000483:Alleles; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D019175:DNA Methylation / Q000187:drug effects; D006801:Humans; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods*; D013447:Sulfites / Q000494:pharmacology
chemical_list: D013447:Sulfites; D004247:DNA; C009279:sodium bisulfite