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 one of NEB's enzymes that are Time-Saver qualified. 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.
- Do degenerate recognition sites need to be palindromic?
- How can I generate a restriction enzyme site map for my sequence?
- How can I search for a restriction enzyme by sequence, overhang or name?
- How should I stop my restriction digest?
- How stable is a particular restriction enzyme?
- I don't see any cleavage after my restriction digest. What factors can interfere with cleavage?
- Is extended digestion (incubation times > 1 hour) recommended?
- What does it mean to be Time-Saver™ qualified?
- What information is available in the Restriction Enzyme Database (REBASE)?
- When is star activity a concern?
- When should I choose the HF version of the enzyme?
- How should I set up a restriction digest?
- What does HF® refer to following the name of a restriction enzyme?
Restriction Enzymes at NEB: Over 30 years of Innovation
A Modern Day Gene Genie Sir Richard Roberts on Rebase
- DNA Sequences and Maps Tool
- Alphabetized List of Recognition Sequences
- Cleavage Of Supercoiled DNA
- Compatible Cohesive Ends and Generation of New Restriction Sites
- Dam-Dcm and CpG Methylation
- Enzymes with Multiple Recognition Sequences
- Enzymes with Nonpalindromic Sequences
- Frequencies of Restriction Sites
- Interrupted Palindromes
- Recleavable Blunt Ends
- Recleavable Filled-in 5' Overhangs
- Restriction Enzyme Troubleshooting Guide
- Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
- Activity of Restriction Enzymes in PCR Buffers
- Alteration of Apparent Recognition Specificities Using Methylases
- Cleavage Close to the End of DNA Fragments
- Dam and Dcm Methylases of E. coli
- Double Digests
- Heat Inactivation
- NEBuffer Activity/Performance Chart with Restriction Enzymes
- Optimizing Restriction Endonuclease Reactions
- Restriction Enzyme Diluent Buffer Compatibility
- Restriction Enzyme Tips
- Restriction of Foreign DNA by E. coli K-12
- Site Preferences
- Star Activity
- Enzymes powerful enough to digest in 5-15 minutes
- Flexible enough for overnight incubation
- No special formulation
- No added expense
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How will Time-Saver™ qualified enzymes save you time? Find out from an NEB scientist.
Need a protocol to digest quickly and completely? Try this protocol for Time-Saver™ qualified enzymes from NEB.
NEB has engineered HF® enzymes to eliminate star activity. Learn how, and what this means for your digests.
Watch as Rick Morgan, Research Scientist in the Restriction Enzyme Division, describes his passion for discovering and characterizing restriction enzymes from nature.