title: GSM6862215: hsSKM_low_rep1; Homo sapiens; MNase-Seq
library_construction_protocol: High-density (100% confluency) and low-density (50%-70%confluency) cells were cross-linked with 10mM dithiobis(succinimidyl propionate) (DSP) in PBS at room temperature for 30 min, quenched with 1 M Tris-HCl pH=8.0 at room temperature for 15 min and washed with PBS. Fixed cells were lysed with 400ul RIPA-G buffer + protease inhibitor cocktail (PIC ) +RNase to extract DNA-protein complex. Isolated DNA-protein complex was rinsed with 400ul PBS 5 times and then suspended in 400ul PBS. Ater centrifugation, the pellet was redissolved in 100 ul RIPA-T buffer +RNase by sonication (1s/1s,5s). The redissolved DNA-protein complex was digested with 0.5 ul Micrococcal Nuclease ( #10011,CST) at 37°C for 30 mins. After centrifugation, the supernatant was collected and 2 ul Proteinase K #10012 was added to digested bound protein  at 65°C for 2 hours. The DNA was loaded on 1.5% agarose gel, and then purified by E.Z.N.A. R Gel Extraction Kit (Omega_D2500-02; Omega BIO-TEK).The library construction was carried out using TruSeq DNA PCR-free prep kit (20015963, illumina). Library was sequenced by NovaSeq Sequencer. The library construction was carried out using TruSeq DNA PCR-free prep kit (20015963, illumina). Library was sequenced by NovaSeq Sequencer.