title: GSM8316453: Blood_Recipient_young_HSCs_10M_TX_RRBS_rep3; Mus musculus; Bisulfite-Seq
library_construction_protocol: Genomic DNA of blood cells was extracted and incubated (~20 ng) at 50°C for 3 hours and 75°C for 30 min in a 5 μl lysis buffer [20 mM tris-EDTA (Sigma catalog # T9285-100ML), 20 mM KCl (Thermo Scientific, catalog # AM9640G), 0.3% Triton X-100 (Sigma catalog # 93443-100ML), and protease (1 mg/ml; Qiagen catalog # 19155)]. The lysed cells were then subject to Msp I digestion at 37°C for 3 hours and 80°C for 20 min. Following enzymatic digestion, end-repair mix [1 μl Klenow frag exo− (5 U/μl), 0.2 μl of 10× TangoBuf, and 0.8 μl of dNTP mix] was added and incubated at 37°C for 40 min and 75°C for 15 min. For adaptor ligation, the reaction was incubated at 16°C for 30 min, 4°C overnight, and 65°C for 20 min after adding the ligation mix [2.25 μ