Tth DNA Ligase (EN13)

Tth DNA Ligase (EN13)

103,00 631,00 

NAD-dependent recombinant ligase from Thermus thermophilus. The ligation will occur only if the oligonucleotides are perfectly paired to the complementary target DNA and have no gaps between them. There fore, a single-base substitution can be detected. High thermostability allows ligation using high-stringency hybridization conditions. High specificity and stringency permits sensitive detection of SNPs.

EN13, EN13-025, EN13-250

SKU: EN13 Category:

Available Options:

Additional information

Unit

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Product Information

Tth DNA Ligase catalyzes the NAD-dependent formation of phosphodiester bonds between adjacent 3’ hydroxyl and 5’-phosphate termini in double stranded DNA. It is isolated from Escherichia coli strain containing plasmid carrying the Thermus thermophilus DNA ligase gene.

Tth DNA Ligase is stable and active in optimum ligation temperature range of 45–65°C, which is 7–10°C higher than that of T4 DNA ligase. The final reaction ligation temperature is determined by the Tm of the substrates. High ligation temperature eliminates the nonspecific ligation.

Features:

  • NAD-dependent recombinant DNA ligase derived from Thermus thermophilus and over-expressed in E. coli.
  • High thermostability allows ligation using high-stringency hybridization conditions.
  • High specificity and stringency permits sensitive detection of SNPs.
  • The ligation will occur only if oligonucleotides are perfectly paired to the complementary target DNA and have no gaps between them. Therefore, a single-base substitution can be detected.
  • Equivalent of Ampligase® (Epicentre).

Applications:

  • Ligase Chain Reaction (LCR).
  • Ligation-Rolling Circle Amplification (L-RCA).
  • Repeat Expansion Detection (RED).
  • Simultaneous mutagenesis of multiple sites.
  • Other ligation-based detection methods.

Additional information
Up to twenty freeze/thaw cycles will not compromise 10x Tth Ligation Buffer performance.

Unit definition

One unit of Tth DNA Ligase catalyzes the ligation of 50% of the cossites present in 1 µg of bacteriophage lambda DNA in 1 minute at 45°C. 1 U (Unit) of Tth DNA Ligase = 1 Ampligase® Unit = 15 cohesive end units (CEU).

Protocol & Manual

Tth DNA Ligase (EN13) - Manual

Technical Tips

Storage conditions

All components should be stored at -20°C.

Storage Buffer

50% glycerol, 50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 0.1% Triton X-100, 1 mM DTT

10x Tth ligation buffer

200 mM Tris-HCl (pH 8.3), 250 mM KCl, 100 mM MgCl2, 5 mM NAD, 0.1% Triton X-100

Shipping conditions

Shipped on dry or blue ice.

FAQ

Quality & Safety

Quality control

Tth DNA Ligase activity is assayed in a reaction containing 1 μg of bacteriophage lambda DNA digested with Sal I and Sma I (Control DNA), 1x Tth Ligation Buffer and varying amounts of enzyme for 100 minutes at 45°C. Results are assayed by agarose gel electrophoresis. Free of unspecific DNA nucleases contamination.

Tth DNA Ligase (EN13) - MSDS


Tth DNA Ligase (EN13) - CoA 1.2018 L.626272

Tth DNA Ligase (EN13) - CoA 1.2018 L.646073

Literature