title: GSM3756625: H2A_earlyS_ChIP-seq_rep2; Homo sapiens; ChIP-Seq
library_construction_protocol: After the assay, permeabilized cells were treated with 3% formaldehyde in PBS for 5 min at room temperature. Fixed chromatin of permeabilized cells was digested with MNase in ChIP buffer (10 mM Tris-HCl pH 8.0, 200 mM KCl, 1 mM CaCl2, 0.5% NP-40). The supernatant was incubated with mixed with 50 ml of anti-HA-tag mAb-Magnetic Beads at 4°C overnight with rotation. Then, beads were washed three times with ChIP buffer containing 500 mM KCl and TE. After washing beads, beads were resuspended with 100 ml of ChIP elution buffer (50 mM Tris-HCl pH8.0, 10 mM EDTA and 1% SDS) containing 25 mM NaCl and incubated at 65°C for overnight to reverse the cross-links. Then, DNA was eluted with 0.4 mg/ml Proteinase K. The eluted DNA samples were further purified with a NucleoSpin Gel and PCR clean-up kit (MACHEREY-NAGEL). Libraries were prepared using SMARTer ThruPLEX Tag-seq Kit (Takara Bio).
attributes.tag: GEO Accession
attributes.value: GSM3756625