Alteration of Apparent Recognition Specificities Using Methylases.Activity of Restriction Enzymes in PCR Buffers.Activity at 37☌ for Restriction Enzymes with Alternate Incubation Temperatures.Double Digest Protocol with Standard Restriction Enzymes.Optimizing Restriction Endonuclease Reactions.Structures of the Rare-Cutting Restriction Endonuclease NotI Reveal a Unique Metal Binding Fold Involved in DNA Binding. Lambert, A.R., Sussman, D., Shen, B., Maunus, R., Nix, J., Samuelson, J., Xu, S.Y.Protein Engineering, Design and Selection. Rational design of a chimeric endonuclease targeted to NotI recognition site. Engineering a rare-cutting restriction enzyme: Genetic screening and selection of NotI variants. Samuelson, J.C., Morgan, R.D., Benner, J.S., Claus, T.E., Packard, S.L.
One unit is defined as the amount of enzyme required to digest 1 µg of pBC4 DNA in 1 hour at 37☌ in a total reaction volume of 50 µl. The following reagents are supplied with this product: NEB # This product is related to the following categories: Restriction Endonucleases: N-O, Time-Saver Qualified Restriction Enzymes Products, This product can be used in the following applications: Restriction Enzyme Digestion Reagents Supplied Reagents Supplied coli strain that carries the cloned NotI gene from Nocardia otitidis-caviarum (ATCC 14630). Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design.įor details on NEB’s quality controls for restriction endonucleases, visit our Restriction Enzyme Quality page.Īn E. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 minutes with the flexibility to digest overnight without degradation to DNA. HF enzymes also exhibit dramatically reduced star activity. High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart Buffer single-buffer simplicity means more straightforward and streamlined sample processing. NotI has a High Fidelity version NotI-HF ® ( NEB #R3189). Other Products You May Be Interested In.
Reduce Star Activity with High-Fidelity Restriction Enzymes.
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