Application

Used for the removal of nucleic acid from protein samples.

Benzonase nuclease, or endonuclease from Serratia marcescens, can be used to degrade all forms of DNA and RNA while having no proteolytic activity. Benzonase nuclease can also be used to prepare proteins in microcalorimetric experiments.

The enzyme from Sigma has been used to limit cell clumping during the preparation of chimeric cell mixtures. It has also been used for the preparation of nuclear extracts by digesting DNA and releasing nuclear proteins intimately associated with DNA.

Biochem/physiol Actions

Benzonase^® is a genetically engineered endonuclease from Serratia marcescens. The protein is a dimer of 30 kDa subunits with two essential disulfide bonds. This endonuclease attacks and degrades all forms of DNA and RNA (single stranded, double stranded, linear and circular) and is effective over a wide range of operating conditions. The optimum pH for enzyme activity is found to be 8.0-9.2. It completely digests nucleic acids to 5′- monophosphate terminated oligonucleotides 3 to 5 bases in length. This is ideal for removal of nucleic acids from recombinant proteins and for applications where complete digestion of nucleic acids is desirable. It also reduces viscosity in protein extracts and prevents cell clumping. Pre-treatment of a protein sample improves its resolution on 2D gel electrophoresis by eliminating any bound nucleic acids.

Digests native or heat-denatured DNA and RNA.

Legal Information

Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany

Benzonase^® Nuclease is supplied by Merck KGaA, Darmstadt, Germany and/or its affiliates.

Physical form

Solution in 50% glycerol containing 20 mM Tris HCl, pH 8.0, 2 mM MgCl₂, and 20 mM NaCl.

Unit Definition

One unit will digest sonicated salmon sperm DNA to acid-soluble oligonucleotides equivalent to a δA₂₆₀ of 1.0 in 30 min at pH 8.0 at 37 °C (reaction volume 2.625 ml).