GSM5384421: Cut&Run H3K27me3 conditioned control ASO rep2; Mus musculus; OTHER
Frozen brain tissue was resuspended in 1 ml nuclear extraction buffer (20 mM HEPES-KOH pH 7.9, 10 mM KCl, 0.5 mM Spermidine, 0.1 % TritonX-100, 20 % glycerol, protease inhibitor). Single cell separation was performed using a 21 G needle on a 1 ml syringe to shear the tissue through the needle 2 times. After an incubation time of 5 min on ice, extracted nuclei were pelleted for 3 min at 600 g and prepared for DAPI fluorescence-activated cell sorting in order to remove debris and collect 50,000 single nuclei for further processing. Libraries were prepared following strictly the protocol from Janssens and Hernikoff, Version 3, 2019 (https://www.protocols.io/view/cut-amp-run-targeted-in-situ-genome-wide-profiling-zcpf2vn).