Amplification-free whole-genome bisulfite sequencing by post-bisulfite adaptor tagging

Fumihito Miura, Yusuke Enomoto, Ryo Dairiki, Takashi Ito

研究成果: ジャーナルへの寄稿記事

156 引用 (Scopus)

抄録

DNA methylation plays a key role in epigenetic regulation of eukaryotic genomes. Hence the genome-wide distribution of 5-methylcytosine, or the methylome, has been attracting intense attention. In recent years, whole-genome bisulfite sequencing (WGBS) has enabled methylome analysis at single-base resolution. However, WGBS typically requires microgram quantities of DNA as well as global PCR amplification, thereby precluding its application to samples of limited amounts. This is presumably because bisulfite treatment of adaptor-tagged templates, which is inherent to current WGBS methods, leads to substantial DNA fragmentation. To circumvent the bisulfite-induced loss of intact sequencing templates, we conceived an alternative method termed Post-Bisulfite Adaptor Tagging (PBAT) wherein bisulfite treatment precedes adaptor tagging by two rounds of random primer extension. The PBAT method can generate a substantial number of unamplified reads from as little as subnanogram quantities of DNA. It requires only 100 ng of DNA for amplification-free WGBS of mammalian genomes. Thus, the PBAT method will enable various novel applications that would not otherwise be possible, thereby contributing to the rapidly growing field of epigenomics.

元の言語英語
ページ(範囲)e136
ジャーナルNucleic acids research
40
発行部数17
DOI
出版物ステータス出版済み - 9 1 2012

Fingerprint

Genome
Epigenomics
DNA
hydrogen sulfite
5-Methylcytosine
DNA Fragmentation
DNA Methylation
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Genetics

これを引用

Amplification-free whole-genome bisulfite sequencing by post-bisulfite adaptor tagging. / Miura, Fumihito; Enomoto, Yusuke; Dairiki, Ryo; Ito, Takashi.

:: Nucleic acids research, 巻 40, 番号 17, 01.09.2012, p. e136.

研究成果: ジャーナルへの寄稿記事

@article{b6ebc901da3841f4b285bdd69826f631,
title = "Amplification-free whole-genome bisulfite sequencing by post-bisulfite adaptor tagging",
abstract = "DNA methylation plays a key role in epigenetic regulation of eukaryotic genomes. Hence the genome-wide distribution of 5-methylcytosine, or the methylome, has been attracting intense attention. In recent years, whole-genome bisulfite sequencing (WGBS) has enabled methylome analysis at single-base resolution. However, WGBS typically requires microgram quantities of DNA as well as global PCR amplification, thereby precluding its application to samples of limited amounts. This is presumably because bisulfite treatment of adaptor-tagged templates, which is inherent to current WGBS methods, leads to substantial DNA fragmentation. To circumvent the bisulfite-induced loss of intact sequencing templates, we conceived an alternative method termed Post-Bisulfite Adaptor Tagging (PBAT) wherein bisulfite treatment precedes adaptor tagging by two rounds of random primer extension. The PBAT method can generate a substantial number of unamplified reads from as little as subnanogram quantities of DNA. It requires only 100 ng of DNA for amplification-free WGBS of mammalian genomes. Thus, the PBAT method will enable various novel applications that would not otherwise be possible, thereby contributing to the rapidly growing field of epigenomics.",
author = "Fumihito Miura and Yusuke Enomoto and Ryo Dairiki and Takashi Ito",
year = "2012",
month = "9",
day = "1",
doi = "10.1093/nar/gks454",
language = "English",
volume = "40",
pages = "e136",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "17",

}

TY - JOUR

T1 - Amplification-free whole-genome bisulfite sequencing by post-bisulfite adaptor tagging

AU - Miura, Fumihito

AU - Enomoto, Yusuke

AU - Dairiki, Ryo

AU - Ito, Takashi

PY - 2012/9/1

Y1 - 2012/9/1

N2 - DNA methylation plays a key role in epigenetic regulation of eukaryotic genomes. Hence the genome-wide distribution of 5-methylcytosine, or the methylome, has been attracting intense attention. In recent years, whole-genome bisulfite sequencing (WGBS) has enabled methylome analysis at single-base resolution. However, WGBS typically requires microgram quantities of DNA as well as global PCR amplification, thereby precluding its application to samples of limited amounts. This is presumably because bisulfite treatment of adaptor-tagged templates, which is inherent to current WGBS methods, leads to substantial DNA fragmentation. To circumvent the bisulfite-induced loss of intact sequencing templates, we conceived an alternative method termed Post-Bisulfite Adaptor Tagging (PBAT) wherein bisulfite treatment precedes adaptor tagging by two rounds of random primer extension. The PBAT method can generate a substantial number of unamplified reads from as little as subnanogram quantities of DNA. It requires only 100 ng of DNA for amplification-free WGBS of mammalian genomes. Thus, the PBAT method will enable various novel applications that would not otherwise be possible, thereby contributing to the rapidly growing field of epigenomics.

AB - DNA methylation plays a key role in epigenetic regulation of eukaryotic genomes. Hence the genome-wide distribution of 5-methylcytosine, or the methylome, has been attracting intense attention. In recent years, whole-genome bisulfite sequencing (WGBS) has enabled methylome analysis at single-base resolution. However, WGBS typically requires microgram quantities of DNA as well as global PCR amplification, thereby precluding its application to samples of limited amounts. This is presumably because bisulfite treatment of adaptor-tagged templates, which is inherent to current WGBS methods, leads to substantial DNA fragmentation. To circumvent the bisulfite-induced loss of intact sequencing templates, we conceived an alternative method termed Post-Bisulfite Adaptor Tagging (PBAT) wherein bisulfite treatment precedes adaptor tagging by two rounds of random primer extension. The PBAT method can generate a substantial number of unamplified reads from as little as subnanogram quantities of DNA. It requires only 100 ng of DNA for amplification-free WGBS of mammalian genomes. Thus, the PBAT method will enable various novel applications that would not otherwise be possible, thereby contributing to the rapidly growing field of epigenomics.

UR - http://www.scopus.com/inward/record.url?scp=84866919003&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84866919003&partnerID=8YFLogxK

U2 - 10.1093/nar/gks454

DO - 10.1093/nar/gks454

M3 - Article

C2 - 22649061

AN - SCOPUS:84866919003

VL - 40

SP - e136

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 17

ER -