GSM5501473: E12.5 DFL Del(CBS1) cHiC; Mus musculus; OTHER
Micro-dissected E12.5 proximal and distal forelimb pairs and individual E9.5 trunks were isolated in PBS supplemented with 10% Fetal Calf Serum and dissociated into single cell by collagenase treatment (8ul of collagenase 50mg/ml (Sigma C9697) at 37º for approximately 10min). Samples were cross-linked with 1% formaldehyde (ThermoFisher 28908) for 10 minutes at room temperature, quenched with glycine and washed with PBS containing proteinase inhibitor. After discarding the supernatant cells were stored at -80º. Samples were genotyped by PCR to select for homozygous mutant or control tissues. Further processing was performed as described in (Yakushiji-Kaminatsui et al., 2018) Following the Hi-C protocol, ligated DNA fragments were processed throught the Agilent SureSelectXT protocol to enrich for fragments coming from the probe region chr2:72240000-76840000 (mm9).