GSM7060424: F1 ram sperm, RES, rep2; Ovis aries; Bisulfite-Seq
the sperm were treated with somatic cell lysis buffer (0.1 % SDS, 0.5 % Triton X in DEPC H2O) for 40 min on ice to eliminate somatic cell contamination, followed by centrifugation at 600g for 5 minutes. The sperm pellet was washed and used for DNA extraction using TRIzol reagent. DNA extraction, MspI digestion, end-repair, dA-tailing, adaptor ligation, and bisulfite conversion were conducted in a single tube to minimize DNA loss. Amplified DNA fragments of 200 to 500 bp were size selected using a 2% agarose gel.