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  • EcoRI

    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.

    EcoRI has High Fidelity (HF) and RE-Mix Master Mix versions available.
    cloned at neb recombinant timesaver 5min unique buffer incubation temp heat inactivation cpg blue white
    EcoR-I-cutsite_1
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    R0101S10,000 units20,000 units/ml$55.00Add to Cart
    R0101T10,000 units100,000 units/ml$55.00Add to Cart
    R0101L50,000 units20,000 units/ml$220.00Add to Cart
    R0101M50,000 units100,000 units/ml$220.00Add to Cart
      
    Categories:
    Restriction Endonucleases: C-G,
    Time-Saver™ Qualified Restriction Enzymes
    Applications:
    Restriction Enzyme Digestion

    Description

    EcoRI has a High Fidelity version EcoRI-HF® (NEB #R3101).

    High Fidelity (HF®) Restriction Enzymes have 100% activity in CutSmart™ Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. 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. 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.

    Product Source

    An E. coli strain that carries the cloned EcoRI gene from E. coli RY13 (R.N. Yoshimori).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    NEBuffer EcoRI-2010X

    Properties and Usage

    Unit Definition

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


    Reaction Conditions

    1X NEBuffer EcoRI
    Incubate at 37°C

    1X NEBuffer EcoRI:
    100 mM Tris-HCl
    50 mM NaCl
    10 mM MgCl2
    0.025% Triton® X-100
    pH 7.5 @ 25°C

    Activity in NEBuffers

    NEBuffer 1.1: 25%
    NEBuffer 2.1: 100%
    NEBuffer 3.1: 50%
    CutSmart™ Buffer: 50%

    Diluent Compatibility

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM KPO4
    300 mM NaCl
    0.1 mM EDTA
    200 μg/ml BSA
    50% Glycerol
    10 mM DTT
    0.15% Triton® X-100
    pH 7.5 @ 25°C

    Heat Inactivation

    65°C for 20 min

    Methylation Sensitivity

    dam methylation: Not Sensitive
    dcm methylation: Not Sensitive
    CpG Methylation: Blocked by Some Combinations of Overlapping

    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.
    • Blue-White Screening (Terminal Integrity):
      A sample of DNA vector linearized with a 10-fold excess of a restriction endonuclease, religated and transformed into an E. coli strain expressing the LacZ beta fragment gene results in less than 1% white colonies.
    • Exonuclease Activity (Radioactivity Release):
      The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
    • 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.
    • Non-Specific DNase Activity (16 hour):
      The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.

    Notes

    1. May exhibit star activity in NEBuffer 2.1 or CutSmart Buffer.

    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. What is Star Activity and how can it be avoided?
    2. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
    3. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    4. How can I access the old NEBuffer Activity Chart?
    5. How can I access the old Double Digest Finder?
    6. My restriction enzyme used to work well in the old NEBuffer but the new Performance chart indicates it has lower activity even though the only difference is the addition of BSA and removal of DTT to the new buffers. Why?
    1. Optimizing Restriction Endonuclease Reactions

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