The T4 DNA Ligase MBG 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
- Catalyzes the formation of a phosphodiester bond between juxtaposed 5′-phosphate and 3′-hydroxyl termini
- Exhibits very fast and efficient ligation of DNA fragments with compatible cohesive or blunt ends
Product Details
T4 DNA Ligase MBG is an ATP-dependent recombinant enzyme isolated from Escherichia coli strain used to clone DNA. T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5′-phosphate and 3′-hydroxyl termini in duplex DNA or RNA. It will join both blunt-ended and cohesive-ended restriction fragments of DNA and repair single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.
It is supplied with 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 50 mM KCl, 1 mM DTT, 50% (v/v) glycerol.
One (Weiss) unit of T4 DNA Ligase catalyzes the conversion of 1 nmol of 32P from pyrophosphate into Norit-adsorbable material in 30 minutes at 37°C. One Weiss unit is equivalent to approximately 200 cohesive end units.
Performance
| Assay | Specification |
|---|
| DNase contamination | None detected |
| Exonuclease activity | None detected |
| Endonuclease activity | None detected |
Principle
T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5′-phosphate and 3′-hydroxyl termini in duplex DNA or RNA. It will join both blunt-ended and cohesive-ended restriction fragments of DNA and repair single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.
- The 10x Ligation Buffer and ATP Solution should be thawed and resuspended at room temperature.
- For blunt-end ligations, use higher quantities of both the vector and the insert DNA.
- For sticky cohesive-end ligations, we recommend heating both the vector and the insert DNA before ligation.
- The electrotransformation efficiency may be improved by heat inactivation of the T4 DNA Ligase MBG and purification of the DNA through a spin column purification method.
- We recommend using a 3–10 molar excess of insert DNA over vector DNA.
- The enzyme is inhibited by >200 mM NaCl or KCl concentrations.
- Inactivate the enzyme at 65°C for 10 minutes or at 70°C for 5 minutes.
Procedure
QualityControl
T4 DNA ligase activity is assayed in a reaction containing 1 µg of bacteriophage lambda DNA digested with HindIII, 1x T4 Ligation Buffer and varying amounts of enzyme for 20 minutes at 16°C. Results are assayed by agarose gel electrophoresis. The product is free of unspecific DNA nucleases.
Exonuclease and endonuclease activities were evaluated by gel electrophoresis following incubation of 1 µg of DNA with enzyme in a 50 µL volume for 4 hours at 37°C.
Applications
This is used for applications such as:
- Molecular cloning of PCR products or restriction fragments
- Site-directed mutagenesis
- Nick repair in duplex DNA, RNA or DNA/RNA hybrids
- Self-circularization of linear DNA
- LM PCR methods (Ligation Mediated PCR)
Read more >
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T4 DNA Ligase MBG
T4 DNA Ligase MBG
<p><span style="color: rgb(89, 89, 89); font-size: 14px; letter-spacing: 0.4px; font-family: Tahoma;">The T4 DNA Ligase MBG is intended for molecular biology applications. </span></p><p><span style="color: rgb(89, 89, 89); font-size: 14px; letter-spacing: 0.4px; font-family: Tahoma;">This product is neither intended for the diagnosis, prevention or treatment of a disease, </span></p><p><span style="color: rgb(89, 89, 89); font-size: 14px; letter-spacing: 0.4px; font-family: Tahoma;">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;">Catalyzes the formation of a phosphodiester bond between juxtaposed 5′-phosphate and 3′-hydroxyl termini</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Exhibits very fast and efficient ligation of DNA fragments with compatible cohesive or blunt ends</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><br></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;">T4 DNA Ligase MBG is an ATP-dependent recombinant enzyme isolated from <em style="padding: 0px; border: 0px;">Escherichia coli</em> strain used to clone DNA. T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5′-phosphate and 3′-hydroxyl termini in duplex DNA or RNA. It will join both blunt-ended and cohesive-ended restriction fragments of DNA and repair single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.</p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"><br>It is supplied with 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 50 mM KCl, 1 mM DTT, 50% (v/v) glycerol.</p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"><br>One (Weiss) unit of T4 DNA Ligase catalyzes the conversion of 1 nmol of 32P from pyrophosphate into Norit-adsorbable material in 30 minutes at 37°C. One Weiss unit is equivalent to approximately 200 cohesive end units.</p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"><br></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><br></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: 1241px; padding: 0px; border: 0px; height: 223px;"><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;"><th style="background: rgb(242, 247, 252); width: 618px; padding: 12px 16px; font-weight: 500; font-stretch: normal; border: 0px;">Assay</th><th style="background: rgb(242, 247, 252); width: 618px; padding: 12px 16px; font-weight: 500; font-stretch: normal; border: 0px;">Specification</th></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><br></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;">T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5′-phosphate and 3′-hydroxyl termini in duplex DNA or RNA. It will join both blunt-ended and cohesive-ended restriction fragments of DNA and repair single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.</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;">The 10x Ligation Buffer and ATP Solution should be thawed and resuspended at room temperature.</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">For blunt-end ligations, use higher quantities of both the vector and the insert DNA.</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">For sticky cohesive-end ligations, we recommend heating both the vector and the insert DNA before ligation.</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">The electrotransformation efficiency may be improved by heat inactivation of the T4 DNA Ligase MBG and purification of the DNA through a spin column purification method.</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">We recommend using a 3–10 molar excess of insert DNA over vector DNA.</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">The enzyme is inhibited by >200 mM NaCl or KCl concentrations.</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Inactivate the enzyme at 65°C for 10 minutes or at 70°C for 5 minutes.</li></ul><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"><br></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><br></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;">Quality<span style="padding: 0px; font-weight: 700; border: 0px;"></span>Control</p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">T4 DNA ligase activity is assayed in a reaction containing 1 µg of bacteriophage lambda DNA digested with HindIII, 1x T4 Ligation Buffer and varying amounts of enzyme for 20 minutes at 16°C. Results are assayed by agarose gel electrophoresis. The product is free of unspecific DNA nucleases.</p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"><br>Exonuclease and endonuclease activities were evaluated by gel electrophoresis following incubation of 1 µg of DNA with enzyme in a 50 µL volume for 4 hours at 37°C.</p><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"><br></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><br></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); 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="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;">Molecular cloning of PCR products or restriction fragments</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Site-directed mutagenesis</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Nick repair in duplex DNA, RNA or DNA/RNA hybrids</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Self-circularization of linear DNA</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">LM PCR methods (Ligation Mediated PCR)</li></ul></div></section><p><br></p><p><span style="font-family: Tahoma; font-size: 14px;"> </span></p>
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