dsDNase HL

For the rapid and safe purification of RNA or protein samples from contaminating DNA

S_1358_9_LS_OEM_dsDNase_HL_500_U
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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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$433.00
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Quantity
500 U
2500 U
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.
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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

Quick-Start Protocols (1)
Safety Data Sheets (1)
Certificates of Analysis (1)