GSM5145204: HEK293_siBAHD1_FLAG_mut#1_W667G_cell_PE_CR2; Homo sapiens; OTHER
Cells were trypsinized with 0.05% trypsin and pellets were collected for CUT&RUN. CUT&RUN was performed according to manufacturer's instructions (CUTANA™ pAG-MNase for ChIC/CUT&RUN, EpiCypher, Cat# 15-1116) with the antibodies against FLAG (Sigma cat#F1804). WT-BAHD1-expressing cells labeled as WT#1_SE were processed in parallel with samples labeled as EV (which is a negative control with empty vector) and mut#1_W667G_SE (which expressed a BAHD1 W677G mutant), followed by the single-end (SE) sequencing; the rest of libraries for WT and the two BAH mutants of BAHD1 (WT#2_PE, mut#1_W667G_PE and mut#2_Y669A_PE) were processed in a separate experiment and subjected to paired-end (PE) sequencing later on. 10 ng of the purified CUT&RUN-enriched DNA was used for preparation of multiplexed sequencing libraries with the NEB Ultra II DNA Library Prep Kit per manufacturer's instructions (NEB #E7103).