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 not active against single stranded DNA or RNA and blunt ended DNA.
Available Options:
| SKU | Unit | ||
|---|---|---|---|
 | EN13-250 | 2500 U (5 U/μl eq. 75 CEU/μl) | Ask for an offer |
| EN13-025 | 250 U (5 U/μl eq. 75 CEU/μl) | Ask for an offer |
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 not active against single stranded DNA or RNA and blunt ended DNA. Enzyme 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.
- Equivalent of Ampligase® (Epicentre).
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)
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 cos sites 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).
Tth DNA Ligase (EN13) - Manual
Storage conditions
All components should be stored at -20°C.
Storage Buffer
50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 0.1% Triton X-100, 1 mM DTT, 50% glycerol
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.
Quality control
Tth DNA Ligase activity is tested in reaction with bacteriophage lambda DNA digested with SalI and SmaI, with a dilution series of ligase. Results are assayed by agarose gel electrophoresis. Free of unspecific DNA and RNA nucleases contamination.
Tth DNA Ligase (EN13) - MSDS
Tth DNA Ligase (EN13) - L. 646073 for Protocol v. 1.2017
Tth DNA Ligase (EN13) - L. LCE675875
Tth DNA Ligase (EN13) - L. LCE745774
Tth DNA Ligase (EN13) - L. LCZ785875
Tth DNA Ligase (EN13) - L. LCE646073
Tth DNA Ligase (EN13) - L. LCZ616174
-
Circle-to-circle amplification coupled with microfluidic affinity chromatography enrichment for in vitro molecular diagnostics of Zika fever and analysis of anti-flaviviral drug efficacy
Authors Ruben R.G. Soares, Aleksandra Pettke, Agustín Robles-Remacho, Sahar Zeebaree, Sibel Ciftci, Marianna Tampere, Aman Russom, Marjo-Riitta Puumalainen, Mats Nilsson, Narayanan Madaboosi Products phi29 Polymerase (EN20), Tth DNA Ligase (EN13) Year 2021 Source Sensors and Actuators B: Chemical (336) -
In Situ Sequencing: A High-Throughput, Multi-Targeted Gene Expression Profiling Technique for Cell Typing in Tissue Sections
Authors Markus M. Hilscher, Daniel Gyllborg, Chika Yokota, Mats Nilsson Products RIBOPROTECT Hu RNase Inhibitor (RT35), RNase H (RT34), T4 DNA Ligase (EN11), TRANSCRIPTME Reverse Transcriptase (RT32), Tth DNA Ligase (EN13) Year 2020 Source In Situ Hybridization Protocols. Methods in Molecular Biology, vol 2148 -
Sub-attomole detection of HIV-1 using padlock probes and rolling circle amplification combined with microfluidic affinity chromatography
Authors Ruben R.G. Soares, João C. Varela, Ujjwal Neogi, Sibel Ciftci, Manickam Ashokkumar, Inês F. Pinto, Mats Nilsson, Narayanan Madaboosi, Aman Russom, Products BSA (Bovine Serum Albumin) Fraction V (EN17) – To be discontinued, Tth DNA Ligase (EN13) Year 2020 Source Biosensors and Bioelectronics, Volume 166 -
Stable transformation of unicellular green alga Coccomyxa subellipsoidea C-169 via electroporation
Authors Kania, K., Zienkiewicz, M., Drożak, A. Products RNase H (RT34), Tth DNA Ligase (EN13) Year 2020 Source Protoplasma 257, 607–611 -
Hybridization-based in situ sequencing (HybISS) for spatially resolved transcriptomics in human and mouse brain tissue
Authors Daniel Gyllborg, Christoffer Mattsson Langseth, Xiaoyan Qian, Eunkyoung Choi, Sergio Marco Salas, Markus M Hilscher, Ed S Lein, Mats Nilsson Products phi29 Polymerase (EN20), RIBOPROTECT Hu RNase Inhibitor (RT35), TRANSCRIPTME Reverse Transcriptase (RT32), Tth DNA Ligase (EN13) Year 2020 Source Nucleic Acids Research