GSM6948213: CaptureHiC-mm-Wt-FLHL-E115-tecRep1-L11678; Mus musculus; Hi-C
Forelimb and hindlilmb buds were micro-dissected from E11.5 mouse embryos in 1xPBS, a single cell suspension was made using Trypsin, then samples were crosslinked using 2% PFA, snap frozen, and stored at -80 °C. Snap frozen samples were first digested using DpnII, religated using the T4 DNA ligase (Thermo Fisher Scientific). The chimeric chromatin products were then decrosslinked overnight. RNase A treated chromatin was then precipitated.The precipiated chromatin was sheared using a Covaris sonicator (duty cycle: 10%; intensity: 5; cycles per burst: 200; time: 6 cycles of 60 s each; set mode: frequency sweeping; temperature: 4–7 °C). Adaptors were added to the sheared DNA and amplified according to the manufacturer's instructions for Illumina sequencing (Agilent). The library was hybridized to the custom-designed SureSelect beads and indexed for sequencing following the manufacturer's instructions (Agilent). Capture Hi-C