title: GSM8583531: Huh7,OE,input; Homo sapiens; ChIP-Seq
library_construction_protocol: Cells were crosslinked in 1% formaldehyde solution for 10 min followed by reaction termination in 125mM glycine. The cell pellet was washed twice with 1×PBS and then lysed in Tris-EDTA buffer (10 mM Tris, 1 mM EDTA pH8.0) at 4℃ for 15 min. The cell lysate containing nucleic acid-protein complexes was sheared by ultrasonication (Bioruptor™, Diagenode) and then centrifuged to obtain the supernatant, one-tenth of which was aliquoted as input, followed by protein removal by digestion with Proteinase K at 55℃ for 2hr. The libraries were purified by purification beads (Vazyme, N411). Dynabeads protein A magnetic beads were eluted with PBS twice, and subjected to immunoprecipitation by adding 5μg antibody per ChIP conjugated incubated at 4℃ for 16hr. Then, followed by protein removal by digestion with Proteinase K at 55℃ for 2h. The resulting DNA fragments were purified with 1x DNA clean beads and subjected to library construction with VAHTS Universal DNA Library Prep Kit for Illumina (V3，ND607). After immunoprecipitation, beads 1x DNA clean beads were recovered using a magnet and washed DNA was eluted and cross-links reverted at 65°C for 4 hours then purified with OIAGEN Kit (MinElute PCR Purification Kit（50）Cat.No.28004) and analyzed on Agilent 2100 Bioanalyzer (Agilent) for measuring fragment size distribution (around 150 bp). The qualified libraries were sequenced on Illumina Nova Seq 6000.
identifiers.namespace: GEO
identifiers.id: GSM8583531_r1