At NEB, enzyme production is linked to basic research in the cloning and overexpression of restriction-modification systems. This focus allows us to provide extremely pure enzymes at concentrations that deliver more flexibility to your experimental design.
Whether you are quickly screening large numbers of clones or setting up overnight digests, you will benefit from the high quality of our enzymes. Typically, a restriction digest involves the incubation of 1 µl of enzyme with 1 µg of purified DNA in a final volume of 50 µl for 1 hour. However, to speed up the screening process, choose a Time-Saver™ qualified enzyme for 5-15 minute digestion reactions. These enzymes will digest 1 µg of substrate DNA in 5-15 minutes using 1 µl of enzyme under recommended reaction conditions, and can also be used safely in overnight digestions. Unlike other suppliers, there is no special formulation, change in concentration or need to buy more expensive new lines of enzymes to achieve digestion in 5-15 minutes, nor do you have to worry if you incubate too long.
In an effort to provide you with as much information as possible, NEB has tested all of its enzymes on unit assay substrate as well as plasmid substrate. We recommend that this be used as a guide as it is not definitive for all plasmids. Restriction enzymes can often show site preference, presumably determined by the sequence flanking the recognition site. In addition, supercoiled DNA may have varying rates of cleavage. Note that there are some enzymes that can digest DNA in 5-15 minutes, but cannot be incubated overnight. These are not Time-Saver qualified.
Since all of our enzymes are rigorously tested for nuclease contamination, you can also safely set up digests for long periods of time without sample degradation. Only NEB can offer enzymes with power and flexibility — the power to digest in 5-15 minutes and the flexibility to withstand overnight digestions with no loss of substrate.