GSM2572257: cerebellum oxBS-seq rep1; Mus musculus; Bisulfite-Seq
The tissues for the technical replicates are from independent mouse, and at least two replicates are included in different assay as indicated. Genomic DNA (Qiagen, Cat#: 51306) were extracted according to the manufacturers’ instructions. The sequencing libraries were constructed as described with little modifications. Briefly, genomic DNA (0.5 to 1 μg) was end-repaired, A-tailed, and ligated to methylated adaptors following the manufacturer’s instructions. The ligated fragments were then purified using Bio-Rad Micro Bio-Spin P-6 SSC column (SSC buffer). After the purification, the samples were denaturized using 1M NaOH and oxidized using oxidant solution (15mM KRuO4 in 0.05M NaOH). After the purification, the DNA was further directed to bisulfite conversion using the EZ DNA methylation Gold kit (Zymo Research) according to the instruction manual. The library was sequenced using Illumina HiSeq X Ten. Paired reads were mapped uniquely to the reference genome (mm10, UCSC) by Bismark. Efficient conversion of 5hmC to uracil were calculated by spiked 5hmC control from Zymo Research (Cat#: D5405)