GSM8032554: MCF10AT1, Bio Rep 1, Library 2; Homo sapiens; OTHER
Cells were trypsinized and counted. 5M cells at a time were pelleted down and resuspended in PBS. Aliquots of 1M cells were generated, spun down, supernatant was removed, and the 1M cell pellets were flash frozen in liquid nitrogen and sent to Cantata Bio for extraction and library prep. The Micro-C library was prepared using the Dovetail® Micro-C Kit according to the manufacturer's protocol. Briefly, the chromatin was fixed with disuccinimidyl glutarate (DSG) and formaldehyde in the nucleus. The cross-linked chromatin was then digested in situ with micrococcal nuclease (MNase). Following digestion, the cells were lysed with SDS to extract the chromatin fragments and the chromatin fragments were bound to Chromatin Capture Beads. Next, the chromatin ends were repaired and ligated to a biotinylated bridge adapter followed by proximity ligation of adapter-containing ends. After proximity ligation, the crosslinks were reversed, the associated proteins were degraded, and the DNA was purified then converted into a sequencing library using Illumina-compatible adaptors. Biotin-containing fragments were isolated using streptavidin beads prior to PCR amplification.