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- Tth DNA Polymerase
Tth DNA Polymerase
Performance
Taq DNA Ligase is isolated from Escherichia coli strain containing plasmid carrying the Thermus thermophilus DNA ligase gene.
Taq DNA Ligase catalyzes the NAD-dependent formation of phosphodiester bonds between adjacent 3’-hydroxyl and 5’-phosphate termini in double stranded DNA. It is not active against single stranded DNA or RNA and blunt ended DNA.
Taq DNA Ligase has no detectable activity in ligating blunt-ended DNA and has no activity on RNA or RNA:DNA hybrids.
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.
Applications
- Ligase Chain Reaction (LCR)
- Ligase Detection Reaction (LDR)
- Next-Generation DNA Sequencing (NGS)
- Repeat Expansion Detection (RED).
- Rolling Circle Amplification (RCA)
- Proximity Ligation Assay (PLA)
- Simultaneous mutagenesis of multiple sites
- Other ligation-based detection methods
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.
- Half-life is 48 hours at 65°C and greater than 1 hour at 95°C.
- High specificity and stringency permits sensitive detection of SNPs.
Concentration:
5 u/ul (75CEU/ul)
Supplied in:
50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, and 0.1% Triton® X-100.
Supplied With:
10X Reaction Buffer: 200 mM Tris-HCl (pH 8.3), 250 mM KCl, 100 mM MgCl2, 5 mM NAD, and 0.1% Triton® X-100.
Unit Definition:
One unit of Tth DNA Ligase catalyzes the ligation of 50% of the cos sites present in 1 ?g of bacteriophage lambda DNA in 1 minute at 45°C.
1 U (Unit) of Tth DNA Ligase = 15 cohesive end units (CEU).
Note:
Up to twenty freeze/thaw cycles will not compromise 10x Tth Ligation Buffer performance.