FAQ: Are slow sites found in common vectors?

SacII exhibits strong site preference. SacII requires two recognition sites to activate cleavage. Try reducing the reaction volume to 10 times the volume of enzyme added, or adding 30 units/microgram and incubating for 2-3 hours. Adding an oligonucleotide which contains the sequence will help if there is only a single site on the target DNA. (Conrad, M., and Topal, M.D. PNAS USA 86, 9707-9711 (1989), and Pein, C.-D., et al (1991) NAR 19; 5139-5142).