dsDNase

• Active in a broad temperature range (10–80°C) and broad pH range (optimum at pH 6.0–9.0)
• Highly active at elevated salt concentrations and other typical buffer additives
• Requires bivalent cations (Mg²⁺ and Ca²⁺) for maximum activity
• Degrades dsDNA to fragments below 10 nt
• Activity towards dsDNA is at least 1000 times higher than towards ssDNA

dsDNase is a 42.8 kDa recombinant endonuclease derived from marine amphipods expressed in Pichia pastoris. The enzyme displays high specific activity towards double-stranded DNA, leaving single-stranded DNA or RNA undamaged in standard conditions. dsDNase is highly active in a broad spectrum of temperatures, buffer conditions and pH.

These features make DNaseMe extremely useful for rapid and RNA-safe degradation of genomic DNA, where the absence of ribonucleases is critical to maintaining the integrity of RNA.

The enzyme hydrolyzes phosphodiester linkages yielding oligonucleotides with a 5′-phosphate and 3′-hydroxyl groups.

It is supplied with 20 mM Tris-HCl, pH 8.0; 50 mM KCl; 5 mM MgCl₂ ; 50% (v/v) glycerol.

One unit (U) 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 and pH 8.0 with herring sperm DNA as a substrate.

The specific activity is similar to bovine DNase I; however, dsDNase is characterized by higher stability in demanding reaction and storage conditions (e.g., high salt and detergent-containing buffers and elevated temperature).

Quality Control 

Protein concentration was determined by SDS-PAGE electrophoresis using Coomassie Blue detection assay.
The presence of RNase activity was determined by gel electrophoresis following incubation of 1 µg of total eukaryotic RNA with 10 U of enzyme in a 20 mL volume for 1 hour at 37°C.
The presence of protease activity was determined by SDS-PAGE with Coomassie Blue detection following incubation of 1 mg 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.

This is used for applications such as:

• Extraction and purification of RNA
• Removal of contaminating genomic DNA from RNA samples
• Degradation of DNA template in transcription reactions
• Reduction of viscosity in biological samples
• Removal of residual DNA during primary stem cell isolation, biopharma and bioprocessing procedures