title: GSM2615739: wagyu 120921; Bos taurus; Bisulfite-Seq
library_construction_protocol: DNA was extracted from longissimus muscle tissues with the DNA extraction kit (Tiangen, Beijing, China) according to the manufacturer’s protocol and concentration of the genomic DNA was determined with a NanoDrop ND-2000 UV spectrophotometer (Thermo Fisher Scientific Inc., USA). 5.2 mg genomic DNA spiked with 26 ng lambda DNA were fragmented to 200-300bp by sonication with a focused-ultra sonicator (Covaris, Woburn, MA, USA), followed by end repair and adenylation. Cytosine-methylated barcodes were ligated to sonicated DNA according to the manufacturer’s protocol. After DNA bisulfite conversion, single-strand DNA fragments were PCR amplified using the KAPA HiFi HotStart Uracil + ReadyMix (2X) (Kapa Biosystems, Wilmington, MA, USA).