title: GSM1263221: Bisulfite-seq_WT-1; Mus musculus; Bisulfite-Seq
library_construction_protocol: Dentate gyri were rapidly micro-dissected bilaterally from adult mice.  Genomic DNA was extracted using the DNeasy Kit (Qiagen). Three µg genomic DNA spiked-in with 0.5% unmethylated lambda DNA (Promega) from each sample was fragmented to ~ 300 bp DNA fragments using Covaris Acoustic System. The ends of fragmented DNA were repaired with T4 DNA polymerase, Klenow DNA polymerase, and T4 PNK were used to convert the overhangs into phosphorylated blunt ends. Klenow Fragment (3’ to 5’ exo minus) was used to add an ‘A’ base to the 3’ end of the blunt phosphorylated DNA fragments. Following ligation of adapters (TruSeq index, 12 indexes), bisulfite conversion was performed using an EZ DNA Methylation-Direct kit (Zymo). PCR was then used to enrich bisulfite-converted DNA fragments to obtain the DNA library suitable for HiSeq2000 sequencing.
attributes.tag: GEO Accession
attributes.value: GSM1263221