The Tth DNA Ligase is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.
Features
- Exhibits high thermostability that allows ligation using high-stringency hybridization conditions
- Possesses high specificity and stringency that permit the sensitive detection of SNPs
Product Details
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. The enzyme is isolated from the Escherichia coli strain containing a plasmid carrying the Thermus thermophilus DNA ligase gene.
It is supplied with 50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 0.1% Triton X-100, 1 mM DTT, 50% glycerol.
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. One unit of Tth DNA Ligaseiq equivalent to 15 cohesive end units (CEU).
Performance
| Assay | Specification |
| Ligase activity | Passed |
| DNase contamination | None detected |
| Exonuclease activity | None detected |
| Endonuclease activity | None detected |
Principle
Tth DNA Ligase is stable and active in an optimum ligation temperature range of 45–65°C, which is 7–10°C higher than T4 DNA ligase. The Tm of the substrates determines the final reaction ligation temperature. High ligation temperature eliminates nonspecific ligation.
Procedure
Quality Control
Tth DNA ligase activity is assayed in a reaction containing 1 µg of bacteriophage lambda DNA digested with SalI and SmaI (Control DNA), 1x Tth Ligation Buffer and varying amounts of enzyme for 100 minutes at 450°C. Results are assayed by agarose gel electrophoresis. The product is free of unspecific DNA nucleases. Results are assayed by agarose gel electrophoresis following incubation of 1 µg of DNA substrate with 5 U of Tth ligase enzyme for 4 hours at 700°C.
Applications
This is used for applications such as:
- 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)
Read more >
126124
EN13
Tth DNA Ligase
<p><span style="color: rgb(89, 89, 89); background-color: rgb(249, 249, 249); font-size: 14px; letter-spacing: 0.4px; font-family: Tahoma;">The Tth DNA Ligase is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.</span></p><h4 _ngcontent-serverapp-c1998474798="" class="section" style="width: calc(66.6667% - 12px); padding: 0px; font-size: 28px; font-family: "Futura W01", Arial, Helvetica, sans-serif; font-stretch: normal; border: 0px; letter-spacing: 0px;"><span style="font-size: 14px;"><span style="font-family: Tahoma;">Features</span></span></h4><p><span style="font-family: Tahoma; font-size: 14px;"><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details></span></p><section _ngcontent-serverapp-c1998474798="" style="padding: 0px; border: 0px;"><wca-lightbox _ngcontent-serverapp-c1998474798="" class="description-box" lg-uid="lg0" style="width: 1236px;"></wca-lightbox><div _ngcontent-serverapp-c1998474798="" wcatablegradient="" class="rich-text product-description" style="padding: 0px; font-size: 18px; border: 0px; letter-spacing: 0.5px;"><ul style="width: calc(66.6667% - 12px); padding: 0px 0px 0px 18px; border: 0px; list-style-image: initial;"><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Exhibits high thermostability that allows ligation using high-stringency hybridization conditions</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Possesses high specificity and stringency that permit the sensitive detection of SNPs</li></ul></div></section><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-navigation="Product Details" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details><p></p><h4 _ngcontent-serverapp-c1998474798="" class="intro" style="width: calc(66.6667% - 12px); padding: 0px; font-size: 40px; border: 0px;"><span style="font-size: 14px;"><span style="font-family: Tahoma;">Product Details</span></span></h4><p><span style="font-family: Tahoma; font-size: 14px;"><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-navigation="Product Details" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details></span></p><section _ngcontent-serverapp-c1998474798="" style="padding: 0px; border: 0px;"><wca-lightbox _ngcontent-serverapp-c1998474798="" class="description-box" lg-uid="lg1" style="width: 1236px;"></wca-lightbox><div _ngcontent-serverapp-c1998474798="" wcatablegradient="" class="rich-text product-description" style="padding: 0px; font-size: 18px; border: 0px; letter-spacing: 0.5px;"><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">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. The enzyme is isolated from the<em style="padding: 0px; border: 0px;"> Escherichia coli</em> strain containing a plasmid carrying the <em style="padding: 0px; border: 0px;">Thermus thermophilus</em> DNA ligase gene.</p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"> </p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">It is supplied with 50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 0.1% Triton X-100, 1 mM DTT, 50% glycerol.</p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"> </p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">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. One unit of Tth DNA Ligaseiq equivalent to 15 cohesive end units (CEU).</p></div></section><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details><p></p><h4 _ngcontent-serverapp-c1998474798="" class="section" style="width: calc(66.6667% - 12px); padding: 0px; font-size: 28px; font-family: "Futura W01", Arial, Helvetica, sans-serif; font-stretch: normal; border: 0px; letter-spacing: 0px;"><span style="font-size: 14px;"><span style="font-family: Tahoma;">Performance</span></span></h4><p><span style="font-family: Tahoma; font-size: 14px;"><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details></span></p><section _ngcontent-serverapp-c1998474798="" style="padding: 0px; border: 0px;"><wca-lightbox _ngcontent-serverapp-c1998474798="" class="description-box" lg-uid="lg2" style="width: 1236px;"></wca-lightbox><div _ngcontent-serverapp-c1998474798="" wcatablegradient="" class="rich-text product-description" style="padding: 0px; font-size: 18px; border: 0px; letter-spacing: 0.5px;"><div style="width: calc(66.6667% - 12px); padding: 0px; border: 0px;"></div><div class="rich-data-table-container" style="padding: 0px; border: 0px;"><div class="rich-data-table" style="padding: 0px; border: 0px;"><table border="1" style="width: 1236px; padding: 0px; border: 0px;"><tbody style="padding: 0px; border: 0px;"><tr style="padding: 0px; border-width: 1px 0px; border-top-style: solid; border-top-color: rgb(188, 188, 187); border-right-style: initial; border-right-color: initial; border-bottom-style: solid; border-bottom-color: rgb(188, 188, 187); border-left-style: initial; border-left-color: initial;"><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">Assay</td><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">Specification</td></tr><tr style="padding: 0px; border-width: 1px 0px; border-top-style: solid; border-top-color: rgb(188, 188, 187); border-right-style: initial; border-right-color: initial; border-bottom-style: solid; border-bottom-color: rgb(188, 188, 187); border-left-style: initial; border-left-color: initial;"><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">Ligase activity</td><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">Passed</td></tr><tr style="padding: 0px; border-width: 1px 0px; border-top-style: solid; border-top-color: rgb(188, 188, 187); border-right-style: initial; border-right-color: initial; border-bottom-style: solid; border-bottom-color: rgb(188, 188, 187); border-left-style: initial; border-left-color: initial;"><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">DNase contamination</td><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">None detected</td></tr><tr style="padding: 0px; border-width: 1px 0px; border-top-style: solid; border-top-color: rgb(188, 188, 187); border-right-style: initial; border-right-color: initial; border-bottom-style: solid; border-bottom-color: rgb(188, 188, 187); border-left-style: initial; border-left-color: initial;"><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">Exonuclease activity</td><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">None detected</td></tr><tr style="padding: 0px; border-width: 1px 0px; border-top-style: solid; border-top-color: rgb(188, 188, 187); border-right-style: initial; border-right-color: initial; border-bottom-style: solid; border-bottom-color: rgb(188, 188, 187); border-left-style: initial; border-left-color: initial;"><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">Endonuclease activity</td><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">None detected</td></tr></tbody></table></div></div><div style="width: calc(66.6667% - 12px); padding: 0px; border: 0px;"></div></div></section><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details><p></p><h4 _ngcontent-serverapp-c1998474798="" class="section" style="width: calc(66.6667% - 12px); padding: 0px; font-size: 28px; font-family: "Futura W01", Arial, Helvetica, sans-serif; font-stretch: normal; border: 0px; letter-spacing: 0px;"><span style="font-size: 14px;"><span style="font-family: Tahoma;">Principle</span></span></h4><p><span style="font-family: Tahoma; font-size: 14px;"><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details></span></p><section _ngcontent-serverapp-c1998474798="" style="padding: 0px; border: 0px;"><wca-lightbox _ngcontent-serverapp-c1998474798="" class="description-box" lg-uid="lg3" style="width: 1236px;"></wca-lightbox><div _ngcontent-serverapp-c1998474798="" wcatablegradient="" class="rich-text product-description" style="padding: 0px; font-size: 18px; border: 0px; letter-spacing: 0.5px;"><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Tth DNA Ligase is stable and active in an optimum ligation temperature range of 45–65°C, which is 7–10°C higher than T4 DNA ligase. The Tm of the substrates determines the final reaction ligation temperature. High ligation temperature eliminates nonspecific ligation.</p></div></section><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details><p></p><h4 _ngcontent-serverapp-c1998474798="" class="section" style="width: calc(66.6667% - 12px); padding: 0px; font-size: 28px; font-family: "Futura W01", Arial, Helvetica, sans-serif; font-stretch: normal; border: 0px; letter-spacing: 0px;"><span style="font-size: 14px;"><span style="font-family: Tahoma;">Procedure</span></span></h4><p><span style="font-family: Tahoma; font-size: 14px;"><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details></span></p><section _ngcontent-serverapp-c1998474798="" style="padding: 0px; border: 0px;"><wca-lightbox _ngcontent-serverapp-c1998474798="" class="description-box" lg-uid="lg4" style="width: 1236px;"></wca-lightbox><div _ngcontent-serverapp-c1998474798="" wcatablegradient="" class="rich-text product-description" style="padding: 0px; font-size: 18px; border: 0px; letter-spacing: 0.5px;"><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"><span style="padding: 0px; font-weight: 700; border: 0px;">Quality Control</span></p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"> </p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Tth DNA ligase activity is assayed in a reaction containing 1 µg of bacteriophage lambda DNA digested with SalI and SmaI (Control DNA), 1x Tth Ligation Buffer and varying amounts of enzyme for 100 minutes at 450°C. Results are assayed by agarose gel electrophoresis. The product is free of unspecific DNA nucleases. Results are assayed by agarose gel electrophoresis following incubation of 1 µg of DNA substrate with 5 U of Tth ligase enzyme for 4 hours at 700°C.</p></div></section><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details><p></p><h4 _ngcontent-serverapp-c1998474798="" class="section" style="width: calc(66.6667% - 12px); padding: 0px; font-size: 28px; font-family: "Futura W01", Arial, Helvetica, sans-serif; font-stretch: normal; border: 0px; letter-spacing: 0px;"><span style="font-size: 14px;"><span style="font-family: Tahoma;">Applications</span></span></h4><p><span style="font-family: Tahoma; font-size: 14px;"><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); letter-spacing: 0.25px;"><wca-product-description _ngcontent-serverapp-c3050437383="" _nghost-serverapp-c1998474798="" data-onedatalayer-source="Product Description" class="ng-star-inserted"></wca-product-description></wca-product-details></span></p><section _ngcontent-serverapp-c1998474798="" style="padding: 0px; border: 0px;"><wca-lightbox _ngcontent-serverapp-c1998474798="" class="description-box" lg-uid="lg5" style="width: 1236px;"></wca-lightbox><div _ngcontent-serverapp-c1998474798="" wcatablegradient="" class="rich-text product-description" style="padding: 0px; font-size: 18px; border: 0px; letter-spacing: 0.5px;"><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">This is used for applications such as:</p><ul style="width: calc(66.6667% - 12px); padding: 0px 0px 0px 18px; border: 0px; list-style-image: initial;"><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Ligase Chain Reaction (LCR)</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Ligase Detection Reaction (LDR)</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Next-Generation DNA Sequencing (NGS)</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Repeat Expansion Detection (RED)</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Rolling Circle Amplification (RCA)</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Proximity Ligation Assay (PLA)</li></ul></div></section><p></p>
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