Cells (about 1 × 109 cells) were Collected and used the QIAamp DNA Mini Kit (Qiagen, Germany) to extracted DNA according to the manufacturer’s instructions. Then DNA concentration was measured using Qubit® DNA Assay Kit in Qubit 3.0 Fluorometer (Life Technologies, CA, United States) and DNA quality was assessed by 1% agarose gel electrophoresis. Methylation libraries were constructed by using the NEBNext® Enzymatic Methyl-seq Kit (NEB) with 200 ng DNA as input amounts. And referred to the manufacturer’s protocol, we prepped libraries with fragments size of ∼400 bp for each sample (no biological replicates, a total of 3 libraries). High-throughput sequencing was performed using the Illumina sequencing platform, and the sequencing read length was 150 bp paired-end reads.
We got 3.3G of raw data in total. And bismark (Krueger and Andrews, 2011 (link)) was used to identify and extract methylation sites. The R package DSS (Wu et al., 2015 (link)) was used to identify differentially methylated regions of different components, using a threshold of P-value < 0.005.
We got 3.3G of raw data in total. And bismark (Krueger and Andrews, 2011 (link)) was used to identify and extract methylation sites. The R package DSS (Wu et al., 2015 (link)) was used to identify differentially methylated regions of different components, using a threshold of P-value < 0.005.
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