Allele-Specific DNA Methylation Detection by Pyrosequencing®

Lasse Sommer Kristensen; Jens Vilstrup Johansen; Kirsten Grønbæk

doi:10.1007/978-1-4939-2715-9_20

Allele-Specific DNA Methylation Detection by Pyrosequencing®

Lasse Sommer Kristensen, Jens Vilstrup Johansen, Kirsten Grønbæk

Institut for Klinisk Medicin

4 Citationer (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.

Originalsprog	Engelsk
Titel	Pyrosequencing : Methods and Protocols
Redaktører	Ulrich Lehmann, Jörg Tost
Antal sider	19
Vol/bind	1315
Publikationsdato	2015
Sider	271-89
ISBN (Trykt)	978-1-4939-2714-2
ISBN (Elektronisk)	978-1-4939-2715-9
DOI	https://doi.org/10.1007/978-1-4939-2715-9_20
Status	Udgivet - 2015

Navn	Methods in molecular biology (Clifton, N.J.)
ISSN	1064-3745

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10.1007/978-1-4939-2715-9_20

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Allele-Specific DNA Methylation Detection by Pyrosequencing®. / Sommer Kristensen, Lasse; Johansen, Jens Vilstrup ; Grønbæk, Kirsten.

Pyrosequencing: Methods and Protocols. red. / Ulrich Lehmann; Jörg Tost. Bind 1315 2015. s. 271-89 (Methods in molecular biology (Clifton, N.J.)).

Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › peer review

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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({\textregistered}) of methylation-specific PCR (MSP) products including a single nucleotide polymorphism (SNP) within the amplicon.",

keywords = "Alleles, DNA, DNA Methylation, Humans, Polymerase Chain Reaction, Sequence Analysis, DNA, Sulfites",

author = "{Sommer Kristensen}, Lasse and Johansen, {Jens Vilstrup} and Kirsten Gr{\o}nb{\ae}k",

year = "2015",

doi = "10.1007/978-1-4939-2715-9_20",

language = "English",

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series = "Methods in molecular biology (Clifton, N.J.)",

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editor = "Ulrich Lehmann and J{\"o}rg Tost",

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T1 - Allele-Specific DNA Methylation Detection by Pyrosequencing®

AU - Sommer Kristensen, Lasse

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.

KW - Alleles

KW - DNA

KW - DNA Methylation

KW - Humans

KW - Polymerase Chain Reaction

KW - Sequence Analysis, DNA

KW - Sulfites

U2 - 10.1007/978-1-4939-2715-9_20

DO - 10.1007/978-1-4939-2715-9_20

M3 - Book chapter

C2 - 26103906

SN - 978-1-4939-2714-2

VL - 1315

T3 - Methods in molecular biology (Clifton, N.J.)

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EP - 289

BT - Pyrosequencing

A2 - Lehmann, Ulrich

A2 - Tost, Jörg

ER -

Allele-Specific DNA Methylation Detection by Pyrosequencing®

Abstract

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