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
