title: GSM8242147: H3K4me1 H3K27M, rep2; Mus musculus; OTHER
library_construction_protocol: Tagmentated DNA was extracted with a phenol chloroform isoamyl alcohol method and resuspended with 1mM Tris-HCl 0.01mM EDTA. CUT&Tag was performed as previously described in Kaya-Okur (2020). In brief, 100,000 bead-bound cells per sample were permeabilized and incubated with primary antibodies targeting the desired epitopes (anti-K4M (ReMAb Biosciences, 31-1250-00S, RM363), anti-H3K4me3 (Active motif, Cat No.39060), anti-H3K4me1 (LifeTech, 710795), anti-H3K27ac (Activ Motif, Cat No.39133), H3K27me3 (CST, C36B11, #9733). 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. After tagmentation and DNA clean-up, libraries were prepared by PCR amplification with barcoded Illumina-compatible primers. Tapestation 2200 analysis using High Sensitivity D5000 ScreenTape (Agilent) was performed to confirm successful library preparation before sequencing.