Allele-Specific DNA Methylation Detection by Pyrosequencing®

Lasse Sommer Kristensen, Jens Vilstrup Johansen, Kirsten Grønbæk

4 Citations (Scopus)

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.

Original languageEnglish
Title of host publicationPyrosequencing : Methods and Protocols
EditorsUlrich Lehmann, Jörg Tost
Number of pages19
Volume1315
Publication date2015
Pages271-89
ISBN (Print)978-1-4939-2714-2
ISBN (Electronic) 978-1-4939-2715-9
DOIs
Publication statusPublished - 2015
SeriesMethods in molecular biology (Clifton, N.J.)
ISSN1064-3745

Keywords

  • Alleles
  • DNA
  • DNA Methylation
  • Humans
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Sulfites

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