GSM4794076: 651-2_hKO (BS-seq); Mus musculus; Bisulfite-Seq
For tissue samples, tissue was dissected from littermate mice in ice-cold PBS, flash-frozen in liquid nitrogen, and stored at -80°C. DNA was isolated from tissue and cultured cells using the DNeasy Kit. DNA was bisulfite converted and prepared for sequencing using the Tecan Ovation Ultralow Methyl-Seq Kit and the Epitect Bisulfite Kit was used for bisulfite conversion. We used alternate bisulfite conversion cycling conditions ([95°C, 5 min; 60°C, 20 min] x 4 cycles, 20°C hold) to ensure lowest possible bisulfite non-conversion rate. Libraries were either sequenced at a depth of 0.01-0.03x genomic coverage using an Illumina MiSeq 2x150 through the Spike-In Cooperative at Washington University in St. Louis, sequenced at 6.4-7.8x coverage per biological rep with a Illumina HiSeq3000 at the Genome Technology Access Center at Washington University in St. Louis, or with a Illumina NextSeq500 at the Edison Family Center for Genome Sciences & Systems Biology.