title: GSM3097919: TGBS_HRCS_rep2; Mus musculus; Bisulfite-Seq
library_construction_protocol: After treatment of sperm lysis buffer, genomic DNA was extracted from each sperm sample. DNA is purified by RNase and pronaseK treatment, and extracted by PCI separation and EtOH precipitation. Libraries for targeted methylome sequencing were prepared according to the post-bisulfite adaptor tagging (PBAT) protocol (Miura F et al., 2012; Miura F and Ito T, 2015) with slight modifications. Briefly, 300 ng of genomic DNA was fragmented to ~500 bp with Covaris S220 and used for target enrichment with SureSelectXT Mouse Methyl-Seq kit (Agilent). The enriched DNA was subjected to PBAT followed by 5 cycles of PCR enrichment with Kapa Library Amplification Kit. Single-read sequencing (100 cycles) was performed on Illumina HiSeq 2500 using HiSeq SR Rapid Cluster Kit v2 and HiSeq Rapid SBS Kit v2. Two indexed libraries were combined with a high concentration of spike-in PhiX control DNA (20%) and loaded on a single lane of the flow cell. Target BS-seq
attributes.tag: GEO Accession
attributes.value: GSM3097919