Abstract
Background Dynamic epigenetic reprogramming occurs during mammalian germ cell development, whereas the targets of this process including DNA demethylation and de novo methylation remain poorly understood. Here, we examined genome-wide methylation profiles in developing primordial germ cells (PGCs) of mice using high-throughput shotgun sequencing of bisulfite-treated DNA (whole-genome shotgun bisulfite sequencing; WGSBS), which accurately quantifies whole-genome methylation levels at single-base resolution.
Highlights
Dynamic epigenetic reprogramming occurs during mammalian germ cell development, whereas the targets of this process including DNA demethylation and de novo methylation remain poorly understood
Materials and methods Using Illumina sequencing libraries, we scaled down the construction and analysis to nanogram quantities of DNA by generating a new WGSBS library, termed the postbisulfite adapter tagging (PBAT) method
PBAT libraries were generated from 2,000-5,000 primordial germ cells (PGCs) and WGSBS analysis was performed using Illumina HiSeq 2000
Summary
Dynamic epigenetic reprogramming occurs during mammalian germ cell development, whereas the targets of this process including DNA demethylation and de novo methylation remain poorly understood. We examined genome-wide methylation profiles in developing primordial germ cells (PGCs) of mice using high-throughput shotgun sequencing of bisulfite-treated DNA (whole-genome shotgun bisulfite sequencing; WGSBS), which accurately quantifies whole-genome methylation levels at single-base resolution
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