GSM8264949: MOLM13, 5uM DB2115, 12hr, ELF1; Homo sapiens; OTHER
CUT&Tag was performed as follows, 5×10^5 cells per cell line were harvested and lightly fixed with 0.5% formaldehyde for 2 minutes. The cells were bound to Concavalin A-coated beads (Bangs laboratories) and incubated with the primary antibody at 4°C overnight. Samples were then incubated with a secondary antibody (guinea pig α-rabbit (Antibodies Online, ABIN101961)) followed by adding pre-loaded pA-Tn5 adapter complex (generated in house). Tagmentation buffer with magnesium was used to induce transposase fragmentation. And PCR based library preparation was conducted (see below). DNA was extracted by phenol/chloroform/isoamyl alcohol and fragments were amplified with NEBNext HiFi 2x PCR Master mix and universal i5 and barcoded i7 primers for 13 cycles. AMPure XP beads (#A63880) were used for post-PCR clean-up of the libraries. Libraries were subject to 35 bp paired-end sequencing using Illumina NextSeq 500 platform with 35bp paired end reads on high output mode at the Einstein Epigenomics core. Fastq files were generated using Picard tools v2.17.1 with adapter trimming by TrimGalore v0.3.7 and QC assessment using FASTQC v0.11.4.