title: WGBS of mus musculus
design: 30 ICMs (three replicates/group) were washed with PBS and snap frozen in liquid nitrogen. 20 ICMs (three replicates/group) were washed with PBS and snap frozen in liquid nitrogen. Samples were collected and sent to ShenZhen BGI Genomics Co., Ltd for Whole Genome Bisulfite Sequencing (WGBS) library construction and sequencing. The construction of the WGBS library followed a standard protocol. Briefly, First, lyse the samples and add unmethylated Lambda DNA and set up the bisulfite conversion reaction program. Next, digest the single-stranded DNA and capture the Oligo 1 products, followed by the addition of oligo 2 tagging to initiate the reaction. After reaction and products purification, the purified DNAs are amplified and the corresponding library quality control protocol will be chosen based on the product requirements. Subsequently, configure and set up the reaction system and program for circularization. Eventually, Circle DNA molecules are duplicated through rolling cycle amplification, leading to the formation of a DNA nanoball (DNB) that contains multiple copies of the DNA. High-quality DNBs are then loaded onto patterned nanoarrays using the high-intensity DNA nanochip technique and sequenced via combinatorial Probe-Anchor Synthesis (cPAS).