GSM9156020: Sim_plus_2_8_S9, FruM; Drosophila simulans; OTHER
Ten brains were used for each CUT&Tag sequencing library. The CUT&Tag procedure and sequencing library preparation protocol was previously described, with some modifications (Ahmad 2020). After dissection, brains were fixed in 1% paraformaldehyde/1X PBS for 1 minute, then transferred to 1X PBS. Brains were washed two additional times for 5 minutes per wash. The rat polyclonal anti-FruM antibody was used at a 1:50 dilution (Sanders and Arbeitman 2008). The secondary rabbit anti-rat IgG (abcam ab6703) was used at 1:100 dilution. For control libraries, the brains were only incubated in secondary antibody. DNA recovery was performed as previously described (Kaya-Okur et al. 2019). Samples underwent 21 cycles of PCR and were purified by agencourt bead purification before sequencing (Ahmad 2020). CUT&Tag