Concentration: ≥2 U/μL Shelf life: 36 months. Storage temperature: -20°C without a defrost cycle. For long-term storage, place at -80C.
The dsDNase HL 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
- Highly active in a broad temperature range (10–47°C)
- Yields low temperature activity to protect RNA or proteins
- Highly active in typical buffer formulations and has a wide pH range with an optimum of 7.0–8.0
- Requires at least 2 mM Mg2+ ions for optimal activity
- Possesses irreversible inactivation at low temperature (15 minutes at 52°C, 1 mM DTT)
Product Details
dsDNase HL (500 U) is a 43.3 kDa heat-labile recombinant endonuclease derived from a cold water eukaryotic organism expressed in Pichia pastoris. The enzyme displays high specific activity towards double-stranded DNA, leaving single-stranded DNA or RNA undamaged in standard conditions.
It is supplied with 20 mM Tris-HCl, pH 8.0; 20 mM NaCl; 2 mM MgCl2 ; 1 mM CaCl2 ; 50% (v/v) glycerol.
One unit is defined as the amount of enzyme that causes an increase in absorbance at 260 nm of 1.0 in 30 minutes at 37°C in 50 mM Tris-HCl buffer, pH 8.0 (25°C) supplemented with 5 mM MgCl2, 0.1 mg/mL BSA and 0.5 mg/mL herring sperm DNA as a substrate.
Performance
| Assay | Specification |
| Protein Purity | >90% |
| RNase activity | None detected |
| Proteolytic activity | None detected |
Principle
dsDNase HL can be quickly inactivated by heat treatment in moderate temperatures. It is intended for applications where the presence of dsDNA influences experiments’ results in thermo-sensitive applications. It is also beneficial for the rapid and safe purification of RNA or protein samples from contaminating DNA.
The enzyme hydrolyzes phosphodiester linkages yielding oligonucleotides with a 5′-phosphate and 3′-hydroxyl groups.
Procedure
Quality Control
Protein concentration was determined by densitometry of SDS-PAGE electrophoresis using Coomassie Blue detection assay.
The presence of RNase activity was determined after incubation of 5 U dsDNase HL with 1 μg of total eukaryotic RNA in a 20 μL volume for 1 hour at 37°C.
The presence of proteolytic activity was determined by SDS-PAGE with Coomassie Blue detection following incubation of 1 μg of BSA with 100 U of enzyme for 20 hours at 37°C.
A 1.575 mL reaction volume containing herring sperm DNA as a substrate is incubated for 30 minutes at pH 8 at 37°C with an enzyme that degrades DNA to acid-soluble oligonucleotides. One unit of the enzyme causes an increase in absorbance at A260 of 1.0.
Applications
This is used for applications such as:
- dsDNA digestion for plasmid and genomic DNA
- RNA and protein samples rapid purification
- PCR, qPCR master mixes and other diagnostic reagents decontamination
- Degradation of DNA template in transcription reactions
Read more >
125976
EN31
Media/Uploads/EN31-005_copy(1).webp
dsDNase HL
<p style="font-size: 19.0008px;"><span style="color: rgb(89, 89, 89); background-color: rgb(255, 255, 255); font-size: 16px; font-family: Tahoma; letter-spacing: 0.25px;">Concentration: ≥2 U/μL Shelf life: 36 months. Storage temperature: -20°C without a defrost cycle. For long-term storage, place at -80C.</span></p><p style="font-size: 19.0008px;"><span style="color: rgb(89, 89, 89); background-color: rgb(249, 249, 249); font-size: 16px; font-family: Tahoma; letter-spacing: 0.4px;">The dsDNase HL 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><span style="font-size: 19.0008px; font-family: Tahoma;"><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><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-family: Tahoma;">Features</span></h4><span style="font-size: 19.0008px; font-family: Tahoma;"><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"><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;">Highly active in a broad temperature range (10–47°C)</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Yields low temperature activity to protect RNA or proteins</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Highly active in typical buffer formulations and has a wide pH range with an optimum of 7.0–8.0</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Requires at least 2 mM Mg<span style="padding: 0px; font-size: 12px; font-stretch: normal; border: 0px; letter-spacing: 0.4px;">2+</span> ions for optimal activity</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Possesses irreversible inactivation at low temperature (15 minutes at 52°C, 1 mM DTT)</li></ul></div></section></wca-product-description></wca-product-details><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><h4 _ngcontent-serverapp-c1998474798="" class="intro" style="width: calc(66.6667% - 12px); padding: 0px; font-size: 40px; font-family: Assistant, sans-serif; border: 0px;"><span style="font-family: Tahoma;">Product Details</span></h4><span style="font-size: 19.0008px; font-family: Tahoma;"><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"><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;">dsDNase HL (500 U) is a 43.3 kDa heat-labile recombinant endonuclease derived from a cold water eukaryotic organism expressed in <em style="padding: 0px; border: 0px;">Pichia pastoris</em>. The enzyme displays high specific activity towards double-stranded DNA, leaving single-stranded DNA or RNA undamaged in standard conditions.</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 20 mM Tris-HCl, pH 8.0; 20 mM NaCl; 2 mM MgCl<span style="padding: 0px; font-size: 12px; font-stretch: normal; border: 0px; letter-spacing: 0.4px;">2</span> ; 1 mM CaCl<span style="padding: 0px; font-size: 12px; font-stretch: normal; border: 0px; letter-spacing: 0.4px;">2</span> ; 50% (v/v) 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 is defined as the amount of enzyme that causes an increase in absorbance at 260 nm of 1.0 in 30 minutes at 37°C in 50 mM Tris-HCl buffer, pH 8.0 (25°C) supplemented with 5 mM MgCl<span style="padding: 0px; font-size: 12px; font-stretch: normal; border: 0px; letter-spacing: 0.4px;">2</span>, 0.1 mg/mL BSA and 0.5 mg/mL herring sperm DNA as a substrate.</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;">Performance</h4><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"> </p><section _ngcontent-serverapp-c1998474798="" style="padding: 0px; font-size: 16px; font-family: "Futura W01", Arial, Helvetica, sans-serif; border: 0px; letter-spacing: 0.25px;"><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="height: 223px; width: 811px; 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;">Protein Purity</td><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">>90%</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;">RNase 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;">Proteolytic activity</td><td style="width: 618px; padding: 12px 16px; border: 0px; text-align: left;">None detected</td></tr></tbody></table></div></div></div></section><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;">Principle</h4><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="font-size: 16px; font-family: "Futura W01", Arial, Helvetica, sans-serif; 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><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; border: 0px;"><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">dsDNase HL can be quickly inactivated by heat treatment in moderate temperatures. It is intended for applications where the presence of dsDNA influences experiments’ results in thermo-sensitive applications. It is also beneficial for the rapid and safe purification of RNA or protein samples from contaminating DNA.</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;">The enzyme hydrolyzes phosphodiester linkages yielding oligonucleotides with a 5′-phosphate and 3′-hydroxyl groups.</p></div></section><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="font-size: 16px; font-family: "Futura W01", Arial, Helvetica, sans-serif; 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;">Procedure</h4><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="font-size: 16px; font-family: "Futura W01", Arial, Helvetica, sans-serif; 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><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; border: 0px;"><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;">Protein concentration was determined by densitometry of SDS-PAGE electrophoresis using Coomassie Blue detection assay.</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;">The presence of RNase activity was determined after incubation of 5 U dsDNase HL with 1 μg of total eukaryotic RNA in a 20 μL volume for 1 hour at 37°C.</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;">The presence of proteolytic activity was determined by SDS-PAGE with Coomassie Blue detection following incubation of 1 μg of BSA with 100 U of enzyme for 20 hours at 37°C.</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;">A 1.575 mL reaction volume containing herring sperm DNA as a substrate is incubated for 30 minutes at pH 8 at 37°C with an enzyme that degrades DNA to acid-soluble oligonucleotides. One unit of the enzyme causes an increase in absorbance at A260 of 1.0.</p></div></section><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="font-size: 16px; font-family: "Futura W01", Arial, Helvetica, sans-serif; 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;">Applications</h4><p style="color: inherit; width: calc(66.6667% - 12px); padding: 0px; font-size: inherit; border: 0px; letter-spacing: inherit;"><wca-product-details _ngcontent-serverapp-c3223413817="" _nghost-serverapp-c3050437383="" data-onedatalayer-source="Product Details" class="ng-star-inserted" style="font-size: 16px; font-family: "Futura W01", Arial, Helvetica, sans-serif; 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><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; border: 0px;"><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;">dsDNA digestion for plasmid and genomic DNA</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">RNA and protein samples rapid purification</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">PCR, qPCR master mixes and other diagnostic reagents decontamination</li><li style="color: inherit; padding: 8px 0px 0px; font-size: inherit; border: 0px; letter-spacing: inherit;">Degradation of DNA template in transcription reactions</li><li></li></ul></div></section><p> </p></div></section></wca-product-description></wca-product-details></span>
2970.07
2517.01
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