GSM9538136: HiC_WD_SUMO_Replicate1_L4; Drosophila melanogaster; Hi-C
Sixty wing discs per replicate were dissected in Schneider's medium and crosslinked in 2% formaldehyde (HBSS, 20 min at room temperature). Crosslinking was quenched with 3 M Tris-Cl (pH 7.5). Samples were homogenized and nuclei isolated using QIAseq Beads (Qiagen). Chromatin was digested, end-labeled, ligated, and de-crosslinked according to the EpiTech Hi-C Kit (Qiagen) protocol with minor modifications (collagenase treatment omitted). DNA was purified using MinElute® columns (Qiagen), sonicated (Covaris S220), and purified again prior to library preparation. Hi-C libraries were prepared using the EpiTech Hi-C Kit (Qiagen) according to the manufacturer's instructions with minor modifications as described above. Libraries were generated from proximity-ligated DNA fragments and processed for high-throughput paired-end sequencing.