Tagmentation-based whole-genome bisulfite sequencing.
Epigenetic modifications such as carbon 5 methylation of the cytosine base in a CpG dinucleotide context are involved in the onset and progression of human diseases. A comprehensive understanding of the role of genome-wide DNA methylation patterns, the methylome, requires quantitative determination of the methylation states of all CpG sites in a genome. So far, analyses of the complete methylome by whole-genome bisulfite sequencing (WGBS) are rare because of the required large DNA quantities, substantial bioinformatic resources and high sequencing costs. Here we describe a detailed protocol for tagmentation-based WGBS (T-WGBS) and demonstrate its reliability in comparison with conventional WGBS. In T-WGBS, a hyperactive Tn5 transposase fragments the DNA and appends sequencing adapters in a single step. T-WGBS requires not more than 20 ng of input DNA; hence, the protocol allows the comprehensive methylome analysis of limited amounts of DNA isolated from precious biological specimens. The T-WGBS library preparation takes 2 d.
Theoretical Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg, Germany.
Qi
Q
Wang
Lei
L
Gu
Andrew
A
Adey
Bernhard
B
Radlwimmer
Wei
W
Wang
Volker
V
Hovestadt
Marion
M
Bähr
Stephan
S
Wolf
Jay
J
Shendure
Roland
R
Eils
Christoph
C
Plass
Dieter
D
Weichenhan
D000818:Animals; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D023281:Genomics / Q000379:methods*; D006801:Humans; D051379:Mice; D017422:Sequence Analysis, DNA / Q000379:methods*; D013447:Sulfites / Q000737:chemistry
D013447:Sulfites; C042345:hydrogen sulfite