GSM9966573: NPAI_Z1 H3K27ac (CUT&RUN); Mus musculus; OTHER
CUT&RUN was performed using the CUTANA ChIC/CUT&RUN kit (EpiCypher #14-1048) following the manufacturer's protocol. Briefly, 1 x 10^6 neurosphere cells were coupled to Concanavalin A beads, permeabilized with 0.01% digitonin, and incubated overnight at 4C with 0.5 ug of target antibody in antibody buffer (H3K4me3 antibody, SNAP-Certified for CUT&RUN, EpiCypher #13-0041; H3K27ac antibody, SNAP-Certified for CUT&RUN and CUT&Tag, EpiCypher #13-0059). The following day, cells were incubated for 10 min with micrococcal nuclease fused to protein A and protein G (pAG-MNase), which was then activated by the addition of CaCl2. After a 2 h incubation at 4C, the reaction was stopped with stop buffer and E. coli DNA was added as spike-in DNA. DNA was isolated using SPRI beads and quantified with a Qubit fluorometer. Libraries were prepared using the CUTANA CUT&RUN Library Prep Kit following the manufacturer's recommendations. Fragment size distribution was assessed on an Agilent TapeStation system. Multiplexed libraries were sequenced on an AVITI24 sequencer (Element Biosciences) at 150 bp paired-end reads.