title: GSM6706009: CnT WT iPSC K36me2 replicate 2; Mus musculus; OTHER
library_construction_protocol: CUT&Tag was performed as previously described (Kaya-Okur, Janssens, Henikoff, Ahmad, & Henikoff, 2020). In brief, 100,000 bead-bound cells per sample were permeabilized and incubated with primary antibodies targeting the desired epitopes (anti-H3K4me3 (active motif 39060), anti-H3K27ac (Millipore Sigma MABE647), H3K27me3 (Cell Signaling 9733), anti-H3.3K36M (Revmab 31-1085-00-S), Sox2 (Cell Signaling 23064)). After incubation with secondary antibody (Guinea Pig anti-Rabbit IgG, antibodies online ABIN101961) to amplify local abundance of immunoglobulins, protein A/G-tethered transposase (pAG-Tn5, Epicypher) was added and bound in situ at target loci. E. coli DNA spike-in was added for normalization purposes. After tagmentation and DNA clean-up, libraries were prepared by PCR amplification with barcoded Illumina-compatible primers. Tapestation 2200 analysis using High Sensitivity D1000 ScreenTape (Agilent) was performed to confirm successful library preparation before sequencing.