GSM7747570: CRMM1-1-ChIP-Seq-rep2.IP; Homo sapiens; ChIP-Seq
Cells were fixed with 1% formaldehyde and sonicated for 8 min. Sonicated chromatin fragments (150 μl) were diluted 10-fold, and Protein G agarose (60 μl) (Millipore, USA) was added and shaken at 4 °C for 2 h. Then, the mixture was briefly centrifuged at 1000 rpm for 5 min at 4 °C. Anti-H3K18la (PTM-1406) was added to the supernatant and incubated overnight at 4 °C. Protein A and protein G magnetic beads (60 μl) (Millipore, USA) were retained in the supernatant for 6 h to pull down the protein at 4 °C. The DNA was released from the bound chromatin after cross-linking reversal and proteinase K treatment, precipitated, and diluted in 100 μ l of 0.2 M. glycine. Purified DNA fragments were constructed and added to ChIP-seq libraries, amplified, and sequenced on an HiSeq 2500 platform (Illumina).