Research output per year
Research output per year
Dulce B. Vargas-Landin, Jahnvi Pflüger, Trung Viet Nguyen, Ryan Lister
Research output: Chapter in Book/Conference paper › Chapter › peer-review
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.
Original language | English |
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Title of host publication | Methods in Molecular Biology |
Subtitle of host publication | Methods and Protocols |
Place of Publication | New York |
Publisher | Humana Press Inc. |
Pages | 383-390 |
Number of pages | 8 |
ISBN (Electronic) | 9781493977741 |
ISBN (Print) | 9781493977734 |
DOIs | |
Publication status | Published - 2024 |
Name | Methods in Molecular Biology |
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Volume | 2842 |
ISSN (Print) | 1064-3745 |
ISSN (Electronic) | 1940-6029 |
Research output: Chapter in Book/Conference paper › Chapter › peer-review