E1014

Benzonase^® Nuclease

≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution

• Synonym(s): Endonuclease from Serratia marcescens
• CAS Number: 9025-65-4
• Enzyme Commission number: 3.1.30.2 (BRENDA, IUBMB)
• MDL number: MFCD00131010
• UNSPSC Code: 12352204
• NACRES: NA.54

Properties

• biological source: Serratia marcescens
• Quality Level: 200
• recombinant: expressed in E. coli
• Assay: ≥90% (SDS-PAGE)
• form: buffered aqueous glycerol solution
• mol wt: 30 kDa
• concentration: ≥250 units/μL
• application(s): research use
• foreign activity: protease, essentially free
• shipped in: wet ice
• storage temp.: −20°C

Description

General description

Benzonase^® nuclease is a highly efficient and genetically engineered endonuclease that originates from Serratia marcescens[1]. This dimeric protein with two essential disulfide bonds [2] is capable of attacking and degrading all forms of DNA and RNA (single-stranded, double-stranded, linear, and circular) under a wide range of operating conditions. Benzonase^® nuclease is capable of removing nucleic acids and enhancing the purity and quality of protein samples.

Application

Benzonase^® Nuclease has been used: as a component in ice-cold lysis buffer C to digest DNA and  RNA to facilitate the complete release of all nuclear proteins[3] in the immunoprecipitation step to release protein complexes from the nucleoplasm and chromatin[4]as a supplement in RIPA to fractionate SHSY5Y cells for immunoprecipitation[5] to remove residual nucleic acids from the aortic roots in decellularization method[6]

Used for the removal of nucleic acid from protein samples.

Biochem/physiol Actions

Benzonase^® Nuclease can completely digest nucleic acids into 5′-monophosphate terminated oligonucleotides of 3 to 5 bases in length, making it the ideal tool for removing nucleic acids from recombinant proteins and for applications that require complete digestion of nucleic acids. In addition to reducing viscosity in protein extracts and preventing cell clumping, pretreatment of protein samples with Benzonase® nuclease can significantly improve their resolution on 2D gel electrophoresis by eliminating any bound nucleic acids. This versatile enzyme can digest both native or heat-denatured DNA and RNA, with its optimum pH for enzyme activity found to be 8.0-9.2. Benzonase^® nuclease is effective at removing host DNA from microbiome samples. In many cases, microbiome samples (such as saliva or skin) will have a high percentage of host DNA that interferes with downstream results. Our experts show that the reduction of host DNA lowers the cost of sequencing while increasing and improving the data. Experimental data is shown in the technical article - Benzonase^® Nuclease for Microbiome Workflows

Digests native or heat-denatured DNA and RNA.

Features and Benefits

• Host DNA depletion in microbiome samples.

• Effective nucleic acid digestion in a variety of workflows.

• Viscosity reduction during protein extraction.

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).

Physical form

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

Other Notes

2024 CiteAb Award Winner for Supplier Succeeding in Parkinson′s Research

Legal Information

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

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

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable
• Personal Protective Equipment: dust mask type N95 (US), Eyeshields, Gloves