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  • TliI

    • This product was discontinued on 01/01/2012
    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.

    cloned at neb recombinant NEBuffer 3 incubation temp heat inactivation no bsa cpg
    Tli-I-cutsite_1
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    Categories:
    Discontinued Products
    Applications:
    Restriction Enzyme Digestion

    Description

    Product Source

    An E. coli strain that carries the TliI gene from Thermococcus litoralis (H.W. Jannasch).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    NEBuffer 3-2010X
    Purified BSA-2010 mg/ml

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA (HindIII digest) in 1 hour at 75°C in a total reaction volume of 50 µl.

    Reaction Conditions

    1X NEBuffer 3
    Supplement with Purified BSA
    Incubate at 75°C

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

    Usage Concentration

    1X

    Activity in NEBuffers

    NEBuffer 1.1: 10%
    NEBuffer 2.1: 50%
    NEBuffer 3.1: 100%
    CutSmart™ Buffer: 50%

    Diluent Compatibility

    Storage Temperature

    -20°C

    Storage Conditions

    25 mM Tris-HCl
    300 mM NaCl
    1 mM DTT
    0.1 mM EDTA
    200 μg/ml BSA
    50% Glycerol
    pH 7.9 @ 25°C

    Heat Inactivation

    No

    Methylation Sensitivity

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

    Activity at Temperature

    @37°C: 10%

    Notes

    1. TliI is a highly thermostable isoschizomer of XhoI.
    2. Incubations of longer than 2 hours are not recommended because of the 75°C optimal incubation temperature.

    FAQs

    1. Is TliI activity sensitive to dam, dcm or mammalian CpG methylation?
    2. How many base pairs should be added at the end of a PCR primer next to the TliI recognition site to guarantee that TliI will cut properly?
    3. Does TliI have any isoschizomers/neoschizomers?
    4. What is the activity of TliI at 37°C?
    5. Are there any additional recommendations for the use of TliI?

    Protocols

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

    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.

    Selection Tools

    Usage Guidelines & Tips

    Troubleshooting Guides

    Interactive Tools

    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.

    Safety Data Sheet

    The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

    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.