GSM9433360: S2-Grh_100uM_aGrh_IP_A_R2; Drosophila melanogaster; ChIP-Seq
ChIP was performed as previously described (Gibson et al. 2024). 25x106 cells were fixed in 0.8% formaldehyde for 7 min and quenched with 125 mM gylcine. Fixed chromatin was sonicated on a Covaris S220 for five 120s rounds with 60s delay, peak power at 170, a duty factor of 10%, and 200 cycles per burst.  IPs were incubated with anti-Grh or anti-HA antibody at 4˚C overnight. Samples were purified using Dynabeads Protein A for Grh (20 uL; Thermo Fisher Scientific) or M-280 Sheep Anti-Mouse IgG for HA (60 uL; Thermo Fisher Scientific). The beads were washed, and chromatin was eluted. IPs and inputs were treated with 90ug RNaseA for 30 mins at 37˚C, and crosslinks were reversed by adding proteinase K overnight at 65˚C. DNA was isolated by phenol:chloroform extraction and concentrated overnight by ethanol precipitation. Libraries were constructed using the NEB Next Ultra II kit.