GSM8943410: ND, Splenic CD8+ T cells, 7 weeks, WGBS; Mus musculus; Bisulfite-Seq
The qualified genomic DNA from splenic CD8+ T cells was used to construct libraries. Briefly, 3 μg DNA spiked with 26 ng unmethylated lambda DNA was fragmented by sonication to produce 200-300 bp fragments with the Covaris S220 (Covaris, MA). Next, the end repair and adenylation reaction were performed. Different cytosine methylated barcodes were ligated to DNA fragments and treated twice with bisulfite using the EZ DNA Methylation-GoldTM kit (Zymo Research, CA). The resulting single-strand DNA fragments were amplified by PCR using the KAPA HiFi HotStart Uracil plus ReadyMix (2×). The Qubit 2.0 Fluorometer was used to quantify the concentration of DNA in the library. The insert size of DNA fragments was assessed by the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA). The libraries for one sample were balanced and mixed with other libraries with different barcodes and sequenced on different lanes of a HiSeq X Ten (Illumina, CA) to generate 150 bp paired-end reads by Biotechnology Corporation (Shanghai, China).