Jan Soroczynski @jsoro.bsky.social · 21/01/2026Micro-C is a huge advance, but MNase must be carefully titrated – over-digest and ends won't ligate, under-digest and you lose resolution. I wanted to overcome this technical challenge. 5/ 100
Jan Soroczynski @jsoro.bsky.social · 21/01/2026Enter CAD (DFF40), a metazoan apoptotic nuclease. It has what we needed: reduced base-composition bias vs MNase, strict endonuclease (no chew-back), and ligation-ready ends (blunt 5'-P/3'-OH). CAD is naturally held inactive by its chaperone-inhibitor ICAD. 6/ 100
Jan Soroczynski @jsoro.bsky.social · 21/01/2026We optimized the CAD:ICAD construct for purification from E. coli and orthogonal activation by TEV protease. Result: consistent genome coverage across a 125-fold CAD concentration range. 7/ 100
