title: GSM4832677: WT-H3K4me3-rep1; Mus musculus; ChIP-Seq
library_construction_protocol: STAR ChIP-seq was performed according to a protocol described previously (Zhang et al., 2016). Briefly, samples were directly lysed and fragmented by micrococcal nuclease (MNase) at 37 °C for 5 min before being incubated with the primary antibody overnight with rotation at 4 °C. The next day, 100 μg Dynabeads Protein A (Thermo Fisher Scientific) was added to each sample and incubated for 2–3 h with rotation at 4 °C. The beads were washed five times with 150 μl radioimmunoprecipitation assay buffer (RIPA) and once with 150 μl lithium chloride buffer. For each sample, beads were resuspended with 27 μl deionized H2O and 1 μl 10× Ex-Taq buffer (Takara, RR006B); 1 μl proteinase K (catalog no. 10910000; Roche) was added to each sample and incubated at 55 °C for 90 min to elute DNA from the beads. After incubation at 80 °C for 40 min to inactivate the proteinase K, the sample was subjected to TELP (tailing, extension, ligation and PCR) library preparation without DNA purification.