NmeAIII

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
R0711S     -20       NmeAIII R0711SVIAL -20 1 x 0.125 ml 2,000 units/ml   rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X

Materials Sold Separately

• rCutSmart™ Buffer

1. Overdigestion with > 5 units of NmeAIII per µg of DNA is not recommended.
2. NmeAIII requires S-adenosylmethionine (SAM) in reaction mixture for optimal activity (SAM is supplied in the enzyme formulation).
3. NmeAIII requires two copies of its recognition sequence for cleavage to occur. Thus, the single NmeAIII site in pUC19 is resistant to cleavage. A 10-fold overdigestion cuts less than half of the DNA.
4. The cleavage point may shift one base pair depending on the DNA sequence context between the recognition site and the position of cleavage. For a given sequence, generally one cut site will predominate.
5. Significant cleavage occurs on ice and at 25°C. 
6. NmeAIII produces a stable partial digestion pattern even with excess enzyme. 1 unit is defined as the amount of enzyme required to produce this stable partial digestion pattern.
7. Based on the stability of the enzyme in the reaction, incubations longer than 1 hr will not result in improved digestion, unless additional enzyme is added. Please refer to Restriction endonuclease survival in a reaction for more information regarding this topic.
8. Requires two or more sites for cleavage. Please review the enzyme list and recommendations in the following table as well as the background information article.
9. Not sensitive to CpG, dcm, or dam methylation.

1. Optimizing Restriction Endonuclease Reactions
2. Restriction Digest Protocol
3. Double Digest Protocol with Standard Restriction Enzymes

• Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
• Activity of Restriction Enzymes in PCR Buffers
• Cleavage Close to the End of DNA Fragments
• Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
• Double Digests
• Heat Inactivation
• NEBuffer Activity/Performance Chart with Restriction Enzymes
• Optimizing Restriction Endonuclease Reactions
• Restriction Endonucleases - Survival in a Reaction
• Restriction Enzyme Diluent Buffer Compatibility
• Restriction Enzyme Tips
• Restriction enzymes requiring multi-sites for efficient cleavage
• Single Letter Codes
• Star Activity
• Traditional Cloning Quick Guide

• 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 Nonpalindromic Sequences
• Frequencies of Restriction Sites
• Isoelectric Points (pI) for Restriction Enzymes
• Isoschizomers
• NEB Diluent and Buffer Table
• Time-Saver™ Qualified Enzymes
• Type IIS Restriction Enzymes
• Why Choose Recombinant Enzymes?

• Competitor Cross-Reference Tool
• DNA Sequences and Maps Tool

• Double Digest Finder
• Enzyme Finder
• NEBcutter™ v3.0
• NEBioCalculator®
• REBASE®

1. How can I search for a restriction enzyme by sequence, overhang or name?
2. How should I stop my restriction digest?
3. How stable is a particular restriction enzyme?
4. When should I choose the HF version of an enzyme?
5. When is star activity a concern?
6. What does it mean to be Time-Saver™ qualified?
7. How should I set up a restriction digest?
8. I don't see any cleavage after my restriction digest. What factors can interfere with cleavage?
9. How can I generate a restriction enzyme site map for my sequence?
10. What information is available in the Restriction Enzyme Database (REBASE)?
11. Is extended digestion (incubation times > 1 hour) recommended?
12. Do degenerate recognition sites need to be palindromic?
13. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
14. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
15. Does NmeAIII require SAM for activity?
16. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

• Restriction Enzyme Troubleshooting Guide

• Restriction Enzyme Cleavage: ‘single-site’ enzymes and ‘multi-site’ enzymes

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

• R0711S_L_v2
• R0711S_v3

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

• R0711S_L_v1_0021305
• R0711S_L_v1_0021308
• R0711S_L_v1_0031311
• R0711S_L_v1_0031405
• R0711S_L_v1_0031410
• R0711S_L_v1_0031509
• R0711S_L_v1_0031606
• R0711S_L_v2_0031606
• R0711S_L_v2_0031609
• R0711S_L_v2_0031707
• R0711S_L_v2_0031803
• R0711S_v2_10011902
• R0711S_v2_10025252
• R0711S_v2_10053313
• R0711S_v2_10081249
• R0711S_v2_10087109
• R0711S_v3_10091788
• R0711S_v3_10095460
• R0711S_v3_10132376
• R0711S_v3_10147373
• R0711S_v3_10161047
• R0711S_v3_10171809
• R0711S_v3_10215051
• R0711S_v3_10239633
• R0711S_v3_10269572
• R0711S_v3_10276583
• R0711S_v3_10306410
• R0711S_v3_10315653
• R0711S_v3_10339663

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• NmeAIII
• rCutSmart™ Buffer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.