DNAseMe dsDNase (EN33)

DNaseMe 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. DNaseMe is highly active in a broad spectrum of temperatures, buffer conditions and pH. The specific activity is similar to bovine DNase I however, DNaseMe is characterized by higher stability in demanding reaction and storage conditions (e.g. high salt and detergent containing

buffers, elevated temperature).

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

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

Features and advantages

• Active in a broad temperature range (10 – 80°C).
• Active in a broad pH range (optimum at pH 6.0 – 9.0).
• Highly active at elevated salt concentrations and other typical buffer additives (Table 1), which significantly improves efficiency and yield of various workflows.
• Requires bivalent cations (Mg²⁺ and Ca²⁺) for maximal activity.
• Degrades dsDNA to fragments below 10 nt.
• The activity towards dsDNA is at least 1000 times higher than towards ssDNA.

Applications

• Extraction and purification of RNA (equivalent of DNase I).
• 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.

Unit definition

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.