GSM2358971: Kc167_Hi-C_primary; Drosophila melanogaster; OTHER
Cross-linked chromatin was released by detergent lysis and Dounce homogenization.  Cross-linked proteins were then biotinylated at cysteine residues and the DNA digested with DpnII.  Digested chromatin was bound to streptavidin beads, thoroughly washed to remove uncross-linked DNA, DNA ends filled in with biotin-14-dATP, and free DNA ends ligated together.  DNA-protein cross-links were reversed, DNA purified, and biotinylated nucleotides marking unligated ends removed. DNA sheared to a size of approximately 500 bp.  DNA was end-repaired and adaptor-ligated by following “NEBNext End Prep” and “Adaptor Ligation” in the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs).  The biotinylated DNA was pulled down with streptavidin beads and PCR amplified by following "PCR Amplification" in the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs).
GEO Accession
GSM2358971