TliI

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.
  3. For enzymes that cannot be heat-inactivated, we recommend using a column for cleanup (such as the Monarch® PCR & DNA Cleanup Kit), or running the reaction on an agarose gel and then extracting the DNA (we recommend Monarch Gel Extraction Kit), or performing a phenol/chloroform extraction.

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 Charts

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. Further information regarding NEB product quality can be found here.
  • 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.