Product Class: Restriction Endonuclease

EcoO109I
neb31 cloned at NEB recombinant time saver dil_A 37 65 Heat dcm

EcoO109I has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10133985. Learn more.

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 RG^GNC_CY
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
  • Restriction Enzyme Cut Site: RG/GNCCY

    • Catalog # Size Concentration
    R0503S 2000.0 units 20000 units/ml

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    Product Information

    Description

    Product Source

    An E. coli strain that carries the cloned EcoO109I gene from E. coli H709c (I. Orskov).
    This product is related to the following categories:
    Restriction Endonucleases C G Products,
    Time-Saver Qualified Restriction Enzymes Products,
    This product can be used in the following applications:
    Fast Cloning: Accelerate your cloning workflows with reagents from NEB,
    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 λ DNA (Hind III digest) 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: 50%
    NEBuffer™ r2.1: 100%
    NEBuffer™ r3.1: 50%
    rCutSmart™ Buffer: 100%

    Diluent Compatibility

    Storage Buffer

    10 mM Tris-HCl
    1 mM DTT
    0.1 mM EDTA
    200 µg/ml Recombinant Albumin
    50% Glycerol
    50 mM KCl
    pH 7.4 @ 25°C

    Heat Inactivation

    65°C for 20 minutes

    Methylation Sensitivity

    dam methylation: Not Sensitive
    dcm methylation: Blocked by Overlapping
    CpG Methylation: Not Sensitive

    Isoschizomers

    DraII+

    Product Notes

    1. Eco0109I is an isoschizomer of DraII.
    2. This enzyme has shown to have lower activity on some supercoiled plasmids, with more than 1 unit required to digest 1 μg plasmid DNA. For complete digestion of 1 μg of plasmid DNA please follow our recommended digestion protocol.
    3. Blocked by overlapping dcm methylation.
    4. star activity may result from a glycerol concentration of >5%

    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. Do degenerate recognition sites need to be palindromic?
    2. 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?
    3. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    4. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
    5. 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.

    Specification Change Notifications

    EcoO109I has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10133985. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin).

    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.

    Trademarks

     

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