title: GSM7729289: SSSavi_mCherry_H3K4me3_ChIP_rep3; Homo sapiens; ChIP-Seq
library_construction_protocol: ChIP-seq: Chromatin from FACS sorted samples were sheared (Covaris, M220) for 5 min (20% duty cycle, 75 peak power, and 200 cycles per burst at 4 ˚C). For dCas9 binding, cells were incubated with 10 µg of HA antibody (Biolegend, #901502), for histone modifications 2 µg of H3K4me3 (Abcam, #ab8580), H3K9me3 (Abcam, #ab8898), or H3K27me3 (Abcam, #ab6002) antibody was used. Each antibody was conjugated to 65 µLprotein G Dynabeads (Invitrogen, #10003D). After wash steps, DNA was eluted, crosslinks were reversed, and immunoprecipitated DNA was purified by Agencourt AMPure XP beads (Beckman Coulter, #A63880). Libraries were prepared from the ChIP eluate containing 1-40 ng input material per replicate using the ThruPLEX DNA-seq 12S kit (Rubicon Genomics, #R400429). After limited PCR amplification (8-10 cycles), libraries were purified using 1x Agencourt AMPure XP beads, and eluted in a final volume of 20 μL.