title: GSM7604325: SETD1A Frame-shift Mutant clone2, replicate 1, day 70; Homo sapiens; OTHER
library_construction_protocol: Cells were resuspended in 100 µL Dig-wash Buffer containing the appropriate primary antibody. Primary antibody incubation was performed on a nutator platform overnight at 4 °C. Cells were washed 3 times with Dig-wash Buffer followed by incubution with an appropriate secondary antibody at RT for 1 hour. Following 3 washes, cells were incubated with pA-Tn5 adapter complex at RT for 1 hour. Following another 3 washes, cells were resuspended in 200 µL Tagmentation buffer at 37 °C for 1 h and then subjected to the addition of 2.25 µL of 0.5 M EDTA, 2.75 µL of 10% SDS and 0.5 µL of 20 mg/mL Proteinase K to stop tagmentation. Cells were then incubated at 55 °C for 2 hours followed by 85 °C for 20 min to inactivate Proteinase K. Genomic DNA was extracted using the ChIP DNA Clean & Concentrator kit Genomic DNA was subjected to 12-14 cycles of PCR amplification for library construction with barcoded i5 and i7 primers, using 25 µL NEBNext HiFi 2× PCR Master mix (NEB). Libraries were cleaned up using Ampure XP beads (Beckman Counter), then sequenced on Illumina NextSeq 550 sequencer with pair-end sequencing (Read1, 37 bp; Read2, 38 bp)