GSM4387223: HepAD38 with DOX #1 3C-HTGTS; Homo sapiens; OTHER
3C libraries were generated as previously described. Briefly, 10 million cells were cross-linked with 1% (v/v) formaldehyde for 10' at room temperature, followed by quenching with glycine at a final concentration of 125 mM. Cells were lysed in 50 mM Tris-HCl, pH 7.5, containing 150 mM NaCl, 5 mM EDTA, 0.5% NP-40, 1% Triton X-100 and protease inhibitors. Nuclei were digested with 500 units of AluI restriction enzyme at 37℃ overnight, followed by ligation under dilute conditions overnight. Crosslinks were reversed and samples were treated with Proteinase K and RNase A prior to DNA precipitation. The 3C libraries were sonicated for 30 s ON and 60 s OFF for two cycles on a Bioruptor Sonicator. Sonicated DNA was linearly amplified with a biotinylated primer. The biotin-labeled single-stranded DNA products were enriched with streptavidin C1 beads, and 3' ends were ligated with the bridge adaptor containing a 6-nucleotide overhang. The adaptor-ligated products were amplified by a nested primer and an adaptor-complementary primer.
GEO Accession
GSM4387223