GSM9032413: 2218, PU.1+, H3K27ac; Homo sapiens; OTHER
Unfixed tissue was dissociated and nuclei were isolated and permeabilised with 0.01% NP-40 and 0.01% digitonin. Chromatin is targeted using histone modification antibodies and tagmented with nano-Tn5. CUT&Tag libraries were prepared using the Illumina Nextera kit. Extracted DNA underwent linear amplification for 8-10 cycles in a thermocycler. 5 ng of this DNA was tagmented with a Tn5 tagmentase. Tagmented DNA was then amplified for 9-11 cycles. Libraries were quantifed using the Qubit HS DNA Sensitivity Assay and the fragment distribution was determined using the Agilent High Sensitivity DNA kit.