TY - JOUR
T1 - Allele-Specific DNA Methylation Detection by Pyrosequencing®
AU - Kristensen, Lasse Sommer
AU - Johansen, Jens Vilstrup
AU - Grønbæk, Kirsten
PY - 2015
Y1 - 2015
N2 - 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.
AB - 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.
U2 - 10.1007/978-1-4939-2715-9_20
DO - 10.1007/978-1-4939-2715-9_20
M3 - Journal article
C2 - 26103906
SN - 1064-3745
VL - 1315
SP - 271
EP - 289
JO - Methods in molecular biology
JF - Methods in molecular biology
ER -