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E1014

Millipore

Benzonase® Nuclease

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

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Synonym(s):
Endonuclease from Serratia marcescens
CAS Number:
Enzyme Commission number:
MDL number:
NACRES:
NA.54

biological source

Serratia marcescens

Quality Level

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. It 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. Choose Benzonase® nuclease for its capabilities in removing nucleic acids and enhancing the purity and quality of protein samples.

Application

Benzonase nuclease, derived from Serratia marcescens, is an effective endonuclease that can efficiently degrade all types of DNA and RNA without any proteolytic activity. This useful enzyme has various applications in protein extraction, microbiome research, and bioprocessing. Benzonase nuclease from Sigma can be utilized to prevent cell clumping during chimeric cell mixtures preparation and for the extraction of nuclear proteins intimately associated with DNA. It is also an ideal solution for effectively removing nucleic acids from protein samples. Additionally, Benzonase nuclease is an essential tool for microbiome research as it can effectively deplete host DNA in microbiome samples. Use Benzonase nuclease for your protein extraction, microbiome research, and bioprocessing needs.
Used for the removal of nucleic acid from protein samples.

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. 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 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 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.

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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Jos J M Drabbels et al.
Blood, 118(19), e149-e155 (2011-09-21)
Microchimerism is defined by the presence of low levels of nonhost cells in a person. We developed a reliable method for separating viable microchimeric cells from the host environment. For flow cytometric cell sorting, HLA antigens were targeted with human
Janus S Jakobsen et al.
Science advances, 5(7), eaaw4304-eaaw4304 (2019-07-17)
The key myeloid transcription factor (TF), CEBPA, is frequently mutated in acute myeloid leukemia (AML), but the direct molecular effects of this leukemic driver mutation remain elusive. To investigate CEBPA mutant AML, we performed microscale, in vivo chromatin immunoprecipitation sequencing
T K Ball et al.
Gene, 57(2-3), 183-192 (1987-01-01)
We are studying exoproteins of the enteric bacterium Serratia marcescens as a model system for the release of extracellular proteins from the cell. In this work we report the cloning of the gene for a secreted nuclease from S. marcescens
T K Ball et al.
Nucleic acids research, 20(19), 4971-4974 (1992-10-11)
The role of the two disulfide bonds found in the Serratia marcescens nuclease were tested by site directed mutagenesis and were found essential for nuclease activity, although slight residual activity remained. The requirement for disulfide bond formation may play a
An extracellular nuclease from Serratia marcescens. II. Specificity of the enzyme.
M Nestle et al.
The Journal of biological chemistry, 244(19), 5219-5225 (1969-10-10)

Articles

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

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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