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  • HphI

    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 timesaver 5min incubation temp heat inactivation dam
    Hph-I-cutsite_1
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    R0158S1,000 units5,000 units/ml$60.00Add to Cart
    R0158L5,000 units5,000 units/ml$240.00Add to Cart
      
    Categories:
    Restriction Endonucleases: H-M,
    Time-Saver™ Qualified Restriction Enzymes
    Applications:
    Restriction Enzyme Digestion

    Description

    Product Source

    An E. coli strain that carries the HphI gene from Haemophilus parahaemolyticus (ATCC 49700).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    CutSmart Buffer-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 CutSmart™ Buffer
    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: 50%
    NEBuffer 2.1: 50%
    NEBuffer 3.1: 10%
    CutSmart™ Buffer: 100%

    Diluent Compatibility

    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: Blocked by Overlapping
    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.
    • 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. It has been suggested that HphI may cleave at N9/N8 depending on the sequence between the recognition and cleavage sites (Kang, C. and Wu, C.-W. (1987) Nucl. Acids Res. 15, 2279-2294; Cho, S.-H. and Kang, C. (1990) Mol. Cells 1, 81?86.). Incubation of > 12 units for over 4 hours on ΦX174 DNA results in additional cleavage products. This has not yet been shown to occur on other DNAs. Low pH and high glycerol concentration enhance this activity.

    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. Is there variability in the cleavage site of HphI?
    2. Does HphI exhibit star activity?
    3. What is the activity of HphI  at 25°C?
    4. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
    5. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    6. Has the conversion to the new buffer system altered any of the properties of the Restriction Enzyme itself?
    7. How can I access the old NEBuffer Activity Chart?
    8. How can I access the old Double Digest Finder?
    9. 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?
    1. Optimizing Restriction Endonuclease Reactions

    Selection Tools

    Usage Guidelines & Tips

    Interactive Tools