WG_RAD.replicate_38 (CRX2258261)
1 g adipose tissue was pulverized; 37% formaldehyde was added to 4% final for cross-linking. After 30 min at 20-25 C, glycine was added to 0.25 M to quench. The mixture was centrifuged at 1500 x g for 10 min, the pellet lysed and homogenized, then centrifuged at 5000 x g to collect nuclei. Nuclei were permeabilized and DNA digested with 200 U MboI at 37 C for 1 h. Ends were filled with biotinylated nucleotides and ligated in dilute volume. Cross-links were reversed, DNA purified and sheared to 300-500 bp, biotin junctions captured with streptavidin beads, and libraries prepared for Illumina NovaSeq 6160.