GSM5240783: 2hr_CapC_rep1; Homo sapiens; OTHER
Cells were harvested in cold PBS and fixed in 1\%~formaldehyde for 10min. Fixed cells were resuspended in lysis buffer, resuspended in PBS and snap-frozen in liquid nitrogen. Thawed cells were pelleted and resuspended in digested overnight with NlaIII. Digested DNA within nuclei were  ligated with T4 DNA ligase overnight. Nuclei were treated with Proteinase K and RNaseA followed by phenol:choloform purification of DNA. The DNA was sonicated to predominantly 200bp fragments. The sonicated material underwent end-repair, dA-tailing, adapter ligation and PCR addition of indices using the NEBNext DNA library Prep Reagent Set and NEBNext Multiplex Oligos for Illumina as per manufacturers instructions, purifying with AxyPrep beads after each step. Indexed libraries were pooled and mixed with human COT DNA, TS-HE universal oligo,  TS-HE Index oligo. This DNA was dried in a heated vacuum centrifuge and resuspended in 1x~hybridisation buffer and hybridisation component A). This DNA-hybridisation mix was denatured and captured by biotinylated probes. Hybridised DNA was washed, amplified and purified. 2 rounds of hybridization were performed. NEBNext DNA library Prep Reagent Set and NEBNext Multiplex Oligos for Illumina