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E1014

Millipore

Benzonase® Nuclease

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

Synonym(s):

Endonuclease from Serratia marcescens

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About This Item

CAS Number:
9025-65-4
Enzyme Commission number:
3.1.30.2 (BRENDA, IUBMB)
MDL number:
MFCD00131010
UNSPSC Code:
12352204
NACRES:
NA.54

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

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General description

Benzonase® nuclease is a highly efficient and genetically engineered endonuclease that originates from Serratia marcescens. This dimeric protein with two essential disulfide bonds 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 in the immunoprecipitation step to release protein complexes from the nucleoplasm and chromatinas a supplement in RIPA to fractionate SHSY5Y cells for immunoprecipitation to remove residual nucleic acids from the aortic roots in decellularization method
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

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

Regulatory Information

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Certificates of Analysis (COA)

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Sequential fractionation and isolation of subcellular proteins from tissue or cultured cells
Baghirova S, et al.
MethodsX, 2, 440-445 (2015)
The involvement of tau in nucleolar transcription and the stress response
Maina M.B., et al.
Acta Neuropathologica Communications, 6(70) (2018)
P Friedhoff et al.
Protein expression and purification, 5(1), 37-43 (1994-02-01)
Overproduction of the extracellular Serratia marcescens nuclease in Escherichia coli results in aggregation and sequestration of a large amount of the protein in inclusion bodies. Only a relatively small amount is secreted into the medium from which it can be
Miles C Scotcher et al.
PloS one, 4(3), e4924-e4924 (2009-03-18)
Botulism, an often fatal neuroparalytic disease, is caused by botulinum neurotoxins (BoNT) which consist of a family of seven serotypes (A-H) produced by the anaerobic bacterium Clostridium botulinum. BoNT, considered the most potent biological toxin known, is a 150 kDa
DONSON facilitates Cdc45 and GINS chromatin association and is essential for DNA replication initiation
Kingsley G, et al.
Nucleic Acids Research, 51(18), 9748?9763-9748?9763 (2023)
View All Related Papers

Articles

Benzonase® Nuclease for Microbiome Workflows

Benzonase® Nuclease for reducing host DNA in microbiome workflows and enhancing taxa identification.

Benzonase® Nuclease Q&A

This page lists nine frequently asked questions and answers about Benzonase® Nuclease.

Benzonase® Nuclease Q&A

This page lists nine frequently asked questions and answers about Benzonase® Nuclease.

Validation of RNAi Knockdown Using Multiple Reaction Monitoring and Protein-AQUA

The field of proteomics is continually looking for new ways to investigate protein dynamics within complex biological samples. Recently, many researchers have begun to use RNA interference (RNAi) as a method of manipulating protein levels within their samples, but the ability to accurately determine these protein amounts remains a challenge. Fortunately, over the past decade, the field of proteomics has witnessed significant advances in the area of mass spectrometry. These advances, both in instrumentation and methodology, are providing researchers with sensitive assays for both identification and quantification of proteins within complex samples. This discussion will highlight some of these methodologies, namely the use of Multiple Reaction Monitoring (MRM) and Protein-AQUA.

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Related Content

Request for Benzonase® endonuclease Safety Plus

The use of Benzonase® endonuclease can significantly reduce the levels of DNA by more than 100,000-fold while also reducing viscosity and protecting downstream equipment from DNA fouling. However, optimization strategies and DoE are critical when it comes to reducing DNA in your process. Setting up a DoE for your Benzonase® endonuclease application can help you find the optimal operation conditions that deliver the required DNA clearance from your process.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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