Benzonase® Nuclease is a geneticallyengineered endonuclease from Serratiamarcescens. It degrades all forms of DNA andRNA (single stranded, double stranded, linearand circular) while having no proteolyticactivity. It is effective over a wide range ofconditions and possesses an exceptionallyhigh specific activity. The enzyme completelydigests nucleic acids to 5′-monophosphateterminated oligonucleotides 2 to 5 bases inlength (below the hybridization limit), whichis ideal for removal of nucleic acids fromrecombinant proteins, enabling compliancewith FDA guidelines for nucleic acidcontamination. The ability of Benzonase torapidly hydrolyze nucleic acids makes theenzyme an excellent choice for viscosityreduction to reduce processing time andincrease yields of protein. For example, theenzyme is compatible with BugBuster andPopCulture Protein Extraction Reagentsand can therefore be added along with thesereagents to eliminate viscosity and removenucleic acids from E. coli extracts.
The enzyme consists of two subunits of30 kDa each. It is functional betweenpH 6 and 10 and from 0-42°C and requires1-2 mM Mg2+ for activation. The enzyme is also active in the presence of ionic and non-ionicdetergents, reducing agents, PMSF(1 mM), EDTA (1 mM) and urea (relativeactivity depends on specific conditions).Activity is inhibited by >150 mM monovalentcations, >100 mM phosphate, >100 mMammonium sulfate, or >100 mM guanidine HCl.
Benzonase Nuclease is available inultrapure (>99% by SDS-PAGE) and pure(>90%) grades at a standard concentrationof 25-29 U/µl and at a high concentration (HC)of 250 U/µl. Both preparations are free ofdetectable protease and have specific activity> 1 × 106 U/mg protein. The >99% puritygrade is tested for endotoxins and contains<0.25 EU/1000 units. The product is suppliedas a 0.2 µm filtered solution in 50% glycerol.Store at -20°C.
Total endotoxin:<0.25 EU/1,000 units.Purity: >99% by SDS-PAGE.
Used for the removal of nucleic acid from protein samples.
Digests native or heat-denatured DNA and RNA.
Toxicity: Standard Handling (A)
One unit is defined as the amount of enzyme that causes a ΔA₂₆₀ of 1.0 in 30 minutes, which corresponds to complete digestion of 37 µg DNA.
Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany
NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany