Tth DNA Polymerase
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.
- 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
- 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.
5 u/ul (75CEU/ul)
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.
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.
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).
Up to twenty freeze/thaw cycles will not compromise 10x Tth Ligation Buffer performance.