title: GSM4827049: RheMac_WT_Placenta_mCG; Macaca mulatta; Bisulfite-Seq
library_construction_protocol: The extracted DNA was mixed with lambda DNA (1:200) and sonicated into an average of 300 bp fragments using Covaris (M220). Sonicated DNA fragments were added by T4 DNA Ligase Buffer (Thermo Scientific), dNTP (10 mM), ATP (10 mM) and END Repair Enzyme Mix, and incubated at room temperature for 1 hour. Then, DNA was purified and added by Klenow Buffer, dATP and Klenow exo- enzyme and incubated at 37°C for 30 minutes. DNA was then purified and added by Quick Ligase Buffer (NEB), Quick Ligase (NEB) and Methylated Adaptors for Illumina Sequencing (Synthesized with all cytosines methylated) and incubated at room temperature for 3 hours. Then, ligated DNA was purified and subjected to sodium bisulfite conversion using the Zymo EZ DNA Methylation-Direct Kit (Zymo Research) following the manufacturer's instruction. Finally, bisulfite converted fragments were PCR amplified using KAPA HiFi Uracil+ Kit following manufacturer's instruction and the libraries were purified and sequenced.