GSM9132090: Mouse liver, Elovl5, ZT12-Vehicle, replicate 2; Mus musculus; OTHER
Mouse livers were harvested at ZT0 and ZT12, and crosslinked with 2% formaldehyde. Nuclei were isolated using lysis buffer, and the fixed chromatin was digested with DpnII. The resulting restriction fragments were ligated using T4 DNA ligase. Crosslinks were reversed with RNase A and Proteinase K, and 3C DNA was purified by phenol/chloroform extraction and resuspended in TLE buffer. The 3C template was then digested with either NlaIII or Csp6I, followed by a second ligation with T4 DNA ligase. 4C templates were purified using phenol/chloroform extraction and AMPure beads, and resuspended in TLE buffer. For library preparation, 800 ng of 4C template were amplified using Roche's Expand Long Template PCR system with bait-specific inverse primers containing Illumina sequencing adaptors. Final libraries were indexed with Nextera XT primers to introduce sample barcodes.