title: GSM4407703: QRICH1-ChIP-seq in QRICH1 KO; Homo sapiens; ChIP-Seq
library_construction_protocol: For ChIP-seq, scrap the cells and follow the ChIP-seq protocol to extract the enriched gDNA from the IP samples. we fixed the cells with 1% formaldehyde and quenched with glycine. We prepared the cell lysate and sonicated the lysates using sonifier (Branson 450 Ultrasonic 102C). We incubated the lysate with anti-QRICH1 antibody (Thermo), anti-ATF4 antibody (Cell signaling), or anti-normal rabbit IgG (Cell signaling) overnight in the cold room. We added the Protein G dynabeads (Invitrogen) for 4 hrs and performed the subsequent ten washes with RIPA bufferbuffer (0.1% SDS, 0.1% DOC, 1% Triton X-100, 1 mM EDTA, 10 mM Tris-HCl pH 8.1, 150 mM NaCl), high-salt RIPA buffer (RIPA with 500 mM NaCl), LiCl buffer (10 mM Tris-HCl pH 8.1, 250 mM LiCl, 0.5% Triton X-100, 0.5% DOC) and PBS. We incubated the DNA overnight at 65 °C to reverse crosslinking and extracted the enriched gDNA fragments after RNase A and Proteinase K treatment.  We generated the DNA library using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina according to the manufacturer's protocol.
attributes.tag: GEO Accession
attributes.value: GSM4407703