GSM5034644: E12.5 inv2 proximal forelimb CR_CTCF; Mus musculus; OTHER
The forelimbs of embryos were dissected and placed into PBS with 10% FCS and 8ul collagenase at 50mg/ml (Sigma C9697) at 37º for approximately 5min. DNA isolated from the CUT&RUN protocol were prepared with KAPA HyperPrep reagents (07962347001) with 2.5ul of adaptors at 0.3uM and ligated for 1 hour at 20ºC. The DNA was amplified for fourteen cycles. Post-amplified DNA was cleaned and size selected using 1:1 ration of DNA:Ampure SPRI beads (A63881) followed by an additional 1:1 wash and size selection with HXB. HXB is equal parts 40% PEG8000 (Fisher FIBBP233) and 5M NaCl. The anti-CTCF  antibody is Active Motif 61311, and the anti-HOXD13 antibody is Abcam Ab19866.
GEO Accession
GSM5034644