Generation of Whole-Genome Bisulfite Sequencing Libraries for Comprehensive DNA Methylome Analysis.

Dulce B Vargas-Landin, Jahnvi Pflüger, Trung Viet Nguyen, Ryan Lister

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Article in Methods in molecular biology (Clifton, N.J.), 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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4 authors.

Dulce B Vargas-LandinAustralian Research Council Centres of Excellence in Plant Energy Biology and Plants for Space, School of Molecular Sciences, The University of Western Australia, Crawley, WA, Australia.

Jahnvi PflügerAustralian Research Council Centres of Excellence in Plant Energy Biology and Plants for Space, School of Molecular Sciences, The University of Western Australia, Crawley, WA, Australia.

Trung Viet NguyenAustralian Research Council Centres of Excellence in Plant Energy Biology and Plants for Space, School of Molecular Sciences, The University of Western Australia, Crawley, WA, Australia.

Ryan ListerAustralian Research Council Centres of Excellence in Plant Energy Biology and Plants for Space, School of Molecular Sciences, The University of Western Australia, Crawley, WA, Australia. ryan.lister@uwa.edu.au.

Funding

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6 · The paper itself

Abstract

Whole-genome bisulfite sequencing (WGBS) enables the detection of DNA methylation at a single base-pair resolution. The treatment of DNA with sodium bisulfite allows the discrimination of methylated and unmethylated cytosines, but the power of this technology can be limited by the input amounts of DNA and the length of DNA fragments due to DNA damage caused by the desulfonation process. Here, we describe a WGBS library preparation protocol that minimizes the loss and damage of DNA, generating high-quality libraries amplified with fewer polymerase chain reaction (PCR) cycles, and hence data with fewer PCR duplicates, from lower amounts of input material. Briefly, genomic DNA is sheared, end-repaired, 3'-adenylated, and ligated to adaptors with fewer clean-up steps in between, minimizing DNA loss. The adapter-ligated DNA is then treated with sodium bisulfite and amplified with a few PCR cycles to reach the yield needed for sequencing.

Indexed as

DNA MethylationPolymerase Chain ReactionSulfitesWhole Genome SequencingCpG IslandsDNAEpigenomeGene LibraryHigh-Throughput Nucleotide SequencingHumansSequence Analysis, DNADNAhydrogen sulfitesodium bisulfiteSulfitesDNA methylationWhole-genome bisulfite sequencing

Identifiers

PMID39012606

DOI10.1016/j.cell.2013.12.019

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