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  • XRN-1

    New size
    recombinant NEBuffer 3 incubation temp heat inactivation
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    M0338S20 units1,000 units/ml$82.00Add to Cart
    M0338L100 units1,000 units/ml$328.00Add to Cart
      
    Categories:
    RNases Products

    Description

    XRN-1 is highly processive 5´→3´ exoribonuclease, requiring 5´ monophosphate. It also acts on 5´ monophosphate ssDNA with greatly reduced efficiency. XRN-1 is not efficient in removing RNAs with recessed 5´P, like tRNA. It does not cleave dsDNA, ssDNA and RNA which contain di, triphosphate, OH and capped 5´ ends.

    Product Source

    Purified from E. coli carrying a plasmid overexpressing the yeast XRN-1 gene (1).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    NEBuffer 3-2010X

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme that digests 1 µg of phosphorylated yeast RNA in 60 minutes at 37°C.

    Reaction Conditions

    1X NEBuffer 3
    Incubate at 37°C

    1X NEBuffer 3:
    100 mM NaCl
    50 mM Tris-HCl
    10 mM MgCl2
    1 mM DTT
    pH 7.9 @ 25°C

    Storage Temperature

    -20°C

    Storage Conditions

    20 mM Tris-HCl
    500 mM NaCl
    2 mM DTT
    0.1 mM EDTA
    50% Glycerol
    0.1% Triton® X-100

    Heat Inactivation

    70°C for 10 min

    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.
    • Endonuclease Activity (Circular Single Stranded DNA):
      The product is tested in a reaction containing circular single stranded DNA. After incubation the percent cleaved is determined by agarose gel electrophoresis.
    • Endonuclease Activity (Nicking):
      The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
    • RNase Activity (1 Hour Digestion):
      The product is tested in a reaction containing a RNA substrate.  After incubation for 1 hour there is no detectable degradation of the RNA substrate as determined by gel electrophoresis.

    Notes

    1. We recommend adding Murine RNase Inhibitor (NEB #M0314) in your reaction at 1 unit/µl to prevent non-specific RNA degradation.

    References

    1. Stevens, A (1980). Biol. Chem. 255, 3080-3085.

    Supporting Documents

    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 the molecular weight of XRN-1?
    2. Is XRN-1 a tagged protein?
    3. Can XRN-1 digest ssDNA containing 5’ monophosphates?
    4. Can XRN-1 digest RNA that contains a di or triphosphate at 5’end?
    5. Is XRN-1 identified with other names in the literature?

    Selection Tools

    We strongly recommend wearing gloves, using nuclease-free tubes and reagents, and thoroughly cleaning pipettes and bench surfaces to avoid RNase contamination.