title: GSM8994897: 4C-CNS11; Mus musculus; OTHER
library_name: GSM8994897
library_construction_protocol: 4C-seq were performed as previously described with slight modifications(Krijger et al., 2020). In brief, 3*106 ILC3s were crosslinked with 1% formaldehyde. Cells were lysed in lysis buffer containing 10mM Tris-HCl (pH8.0), 10mM NaCl, 0.2% Igepal CA630 and protease inhibitors on ice for 50 min. Crosslinked DNA was digested with MboI (NEB) overnight at 37°C, ligated by T4 DNA ligase (NEB), and de-crosslinked with Proteinase K. After DNA purification using the ChIP DNA Clean & Concentrator kit (ZYMO, D5205), a secondary digestion was performed using NlaIII (NEB) at 37°C for 5h, followed by ligation and a final round of DNA purification. The 4C library was amplified using CNS11 specific primers (5'-AGATGGAAATGCTGGTGGAACC-3' and 5'-CCCCTTTGCACCCCTGCATG-3') by Expand Long Template PCR System (Roche, 11681834001).