Product Class: Restriction Endonuclease

NmeAIII
neb31 cloned at NEB recombinant 2site dil_B 37 65 Heat

SAM now included in enzyme formulation – no longer supplied as a separate vial.

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.

NEB restriction endonuclease that recognizes the sequence GCCGAGNNNNNNNNNNNNNNNNNNN_NN^
Isoschizomers | Single Letter Code

Product Introduction

  • 100% activity in rCutSmart™ Buffer (over 210 enzymes are available in the same buffer) allowing for easier double digests

  • Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence

  • 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.

  • Restriction Enzyme Cut Site: GCCGAG(21/19)

Catalog # Size Concentration
R0711S 250.0 units 2000 units/ml

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Product Information

Description

Product Source

An E.coli strain that carries the cloned NmeAIII gene from Neisseria meningitidis 2491 (Achtman, M.)
This product is related to the following categories:
Restriction Endonucleases: N-O Products
This product can be used in the following applications:
Restriction Enzyme Digestion

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of ΦX174 RF I DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X rCutSmart™ Buffer
Incubate at 37°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Activity in NEBuffers

NEBuffer™ r1.1: 10%
NEBuffer™ r2.1: 10%
NEBuffer™ r3.1: <10%
rCutSmart™ Buffer: 100%

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
0.32 mM S-adenosylmethionine (SAM)
500 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 minutes

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive

Product Notes

  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.

Protocols, Manuals & Usage

Protocols

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

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  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?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

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.

Specifications

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]

Certificate Of Analysis

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]

Safety DataSheets

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

Legal and Disclaimers

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 [email protected].

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.

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