GSM4720521: H3K4me1 - Day 0 - GATA6 WT; Homo sapiens; ChIP-Seq
Cells were cross-linked with 1% formaldehyde for 10 minutes at room temperature. The cross-linking reaction was quenched by adding 125mM glycine and incubating at room temperature for 5 minutes. Chromatin was isolated from cell pellets using MNase enzymatic digestion following the manufacturers protocol (Magnetic CHIP kit, ThermoFisher/Pierce). Briefly, cell nuclei were isolated using a membrane lysis buffer. MNase was then added to the isolated nuclei at optimized concentrations and incubated at 37°C for 15 minutes. The enzymatic reaction was stopped with EDTA and digested chromatin was then extracted by brief sonication. Chromatin isolated from 4 individual differentiations were pooled for each condition. Pooled chromatin was incubated with primary antibodies overnight at 4°C. The following day, protein A/G magnetic beads were added to the sample-antibody mixture and incubated at 4°C for 2 hours. Beads were then washed three times in a low salt wash buffer, and once in a high salt wash buffer, before reverse cross-linking in elution buffer. Eluted samples were subjected to proteinase K digestion and the DNA isolated using DNA-binding columns. CHIP DNA samples were submitted to Beijing Genomics Institute (BGI), who performed library preparation and sequencing on the BGISEQ-500 platform.  50bp single end reads were generated and provided as FASTQ files.
GEO
GSM4720521