• My NEB
  • Print
  • PDF
  • Tsp509I

    Tsp509I has been replaced by MluCI

    Discontinued Date

    11/01/2012 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.

    NEBuffer 1 incubation temp heat inactivation no
    Tsp509-I-cutsite_1
    Catalog #SizeConcentrationPriceQtyAdd to Cart
      
    Categories:
    Discontinued Products
    Applications:
    Restriction Enzyme Digestion

    Description

    Product Source

    An E. coli strain that carries the Tsp509I gene from Thermus species (ITI 346).

    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 65°C in a total reaction volume of 50 µl.

    Reaction Conditions

    1X NEBuffer 1
    Incubate at 65°C

    1X NEBuffer 1:
    10 mM Bis-Tris-Propane-HCl
    10 mM MgCl2
    1 mM DTT
    pH 7 @ 25°C

    Usage Concentration

    1%

    Activity in NEBuffers

    NEBuffer 1: 100%
    NEBuffer 2: 100%
    NEBuffer 3: 100%
    NEBuffer 4: NR

    Diluent Compatibility

    Storage Conditions

    10 mM Tris-HCl
    50 mM KCl
    10 mM MgCl2
    1 mM DTT
    0.1 mM EDTA
    200 μg/ml BSA
    50% Glycerol
    pH 7.4 @ 25°C

    Heat Inactivation

    No

    Methylation Sensitivity

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

    Activity at Temperature

    @37°C: 10%

    Notes

    1. Tsp509I is sensitive to methylation by EcoRI Methylase.

    Protocols

    1. Optimizing Restriction Endonuclease Reactions
    2. Double Digest Protocol with Standard Restriction Enzymes

    Selection Tools

    Usage Guidelines & Tips

    Interactive Tools

    Quality Control

    Quality Assurance Statement

    • Tsp509I has been purified free of exonucleases and endonucleases by Finnzymes Oy (Finland), a strategic partner of New England Biolabs, Inc. Quality controls are performed at Finnzymes Oy and confirmed at New England Biolabs, Inc.

    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.

    Material Safety Data Sheets

    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.