GSM8154737: FT-SA11855; Homo sapiens; ChIP-Seq
Four 15-cm dishes of confluent cells were fixed in formaldehyde for 10 min, before quenching the fixation with glycine. Cells were collected, lysed in a sarkosyl-containing lysis buffer and sonicated  to yield 100- to 300-bp genomic DNA fragments. Five micrograms of an antibody raised against H3K4me1 was incubated with blocked magnetic Dynabeads (Life Technologies) at 4 °C for 4 h. Antibody–bead conjugates were incubated with 100 μg of chromatin at 4 °C overnight, with constant agitation. Beads were washed extensively with RIPA buffer and then treated with RNase and proteinase K (both Qiagen). DNA was then eluted from the beads in TE buffer and cleaned up using the QIAquick PCR Purification kit (Qiagen). Sequencing libraries were prepared using a ThruPlex DNA-seq kit (Rubicon R400407), pooled and sequenced on an Illumina HiSeq 5000 for 2 x 150 bp paired-end for an average of 40 million reads (Fulgent Genetics; Temple City, CA).