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dsDNase HL (500 U)
Cat. No. / ID: EN31-005
Concentration: ≥2 U/μL Shelf life: 36 months. Storage temperature: -20°C without a defrost cycle. For long-term storage, place at -80C.
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Quantity
500 U
2500 U
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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.
Need bulk, customized or optimized products for commercial purposes? We also offer support with logistics, compliance and more. Reach out to cooperate with QIAGEN Strategic Partnerships & OEM
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
Resources
Certificates of Analysis (1)