GSM6929349: WGBS Control CD11b+ Biol. Rep. 2, HiSeq 2500; Mus musculus; Bisulfite-Seq
CD11b+ cells were isolated by flow cytometry and DNA was isolated using the PureLink genomic DNA mini kit (Life technologies). DNA was fragmented to an average size of 200 bp using the Adaptive Focused Acoustics Covaris E220 instrument. Before bisulfite treatment, fully unmethylated λ (lambda) DNA (Promega) was spiked into the mouse genomic DNA at a ratio of 1:200. DNA library was prepared by NEBNext® DNA Library Prep Master Mix Set for Illumina (New England BioLabs). Methylated adapters (NeBNext Multiplex Oligos for Illumina, New England BioLabs) were used to determine the bisulfite conversion efficacy. DNA was bisulfite-treated using the MethylCode Bisulfite Conversion Kit (Life Technologies). After the bisulfite conversion the purified DNA was amplified for 4 cycles (low amplification) using Kapa HiFi Uracil+ (Kapa Biosystems). Two independent WGBS samples per genotype were evaluated.