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  • NmeAIII

    This enzyme has transitioned to an improved new buffer system. Visit NEBCutSmart.com for further details.
     
    The new and current Double Digest Finder and current Activity/Performance Chart for the CutSmart buffer system are available. The previous version of the Double Digest Finder, as well as the previous Version of Activity/Performance Chart that use the former buffer system, are still available for your convenience.
    cloned at neb recombinant incubation temp heat inactivation sam
    NmeAIII
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    R0711S250 units2,000 units/ml$65.00Add to Cart
    R0711L1,250 units2,000 units/ml$260.00Add to Cart
      
    Categories:
    Restriction Endonucleases: N-O
    Applications:
    Restriction Enzyme Digestion

    Description

    Product Source

    An E.coli strain that carries the cloned NmeAIII gene from Neisseria meningitidis 2491 (Achtman, M.)

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    CutSmart Buffer-2010X
    S-adenosylmethionine (SAM)-2032 mM

    Properties and 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 CutSmart™ Buffer
    Supplement with 80 μM S-adenosylmethionine (SAM)
    Incubate at 37°C

    1X CutSmart™ Buffer:
    50 mM Potassium Acetate
    20 mM Tris-acetate
    10 mM Magnesium Acetate
    100 μg/ml BSA
    pH 7.9 @ 25°C

    Activity in NEBuffers

    NEBuffer 1.1: 10%
    NEBuffer 2.1: 10%
    NEBuffer 3.1: 10%
    CutSmart™ Buffer: 100%

    Diluent Compatibility

    Storage Temperature

    -20°C

    Storage Conditions

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

    Heat Inactivation

    65°C for 20 min

    Methylation Sensitivity

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

    Quality Control

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Ligation and Recutting (Terminal Integrity):
      After an over-digestion of DNA with a restriction endonuclease the percentage of the DNA fragments ligated with T4 DNA ligase and the percentage that can be recut are determined by agarose gel electrophoresis.

    Notes

    1. Overdigestion with > 5 units of NmeAIII per µg of DNA and incubations > 1 hour are not recommended.
    2. Requires S-adenosylmethionine for optimal activity (supplied with enzyme)
    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.

    Supporting Documents

    Certificate of Analysis

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

    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]

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    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. What are the advantages of using a RE-Mix Restriction Enzyme Master Mix?
    8. How should I set up a restriction digest?
    9. I don't see any cleavage after my restriction digest. What factors can interfere with cleavage?
    10. How can I generate a restriction enzyme site map for my sequence?
    11. What information is available in the Restriction Enzyme Database (REBASE)?
    12. Is extended digestion (incubation times > 1 hour) recommended?
    13. Do degenerate recognition sites need to be palindromic?
    14. My enzyme is no longer Time-Saver™ qualified. What happened?
    15. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
    16. How can I access the old NEBuffer Activity Chart?
    17. How can I access the old Double Digest Finder?
    18. 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?

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