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04716728001 Roche

DNase I recombinant, RNase-free

from bovine pancreas, expressed in Pichia pastoris

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Properties

biological source   bovine pancreas (expressed in Pichia pastoris)
  bovine pancreas (expressed in Pichia pastoris)
form   solution
mol wt   mol wt ~39 kDa
packaging   pkg of 10,000 U
mfr. no.   Roche
optimum pH   7.0-8.0
shipped in   dry ice

Description

Features and Benefits

• Eliminates DNA contamination from any RNA sample
• Contains no detectable RNase or protease activity
• Can be heat inactivated, thereby eliminating the need for organic extraction
• Shipped with an optimized incubation buffer, which supports maximum DNase activity
• Produced via an entirely animal-free process, to eliminate any risks associated with animal-derived material

General description

Recombinant DNase I is a DNA-specific endonuclease.The enzyme catalyzes the degradation of both double- and single-stranded DNA randomly by hydrolyzing phosphodiester linkages to DNA, resulting in a mixture of oligo- and mononucleotides. All material used during the production process of DNase I recombinant is non-animal sourced, resulting in an animal-free product.

Contents
• Recombinant DNase I, RNase-free, 10 U/μl
• Incubation Buffer, 10x concentrated

Other Notes

For life science research only. Not for use in diagnostic procedures.

Packaging

1 kit containing 2 components

Preparation Note

Activator: Bivalent metal ions
Working solution: Storage Buffer: 20 mM Tris-HCl, 50 mM NaCl, 2 mM CaCl2, 2 mM MgCl2, 1 mM dithioerythritol, 0.1 mg/ml Pefabloc SC, 50% glycerol (v/v), pH 7.6 (at 4 °C).
Incubation Buffer (10x): 400 mM Tris-HCl, 100 mM NaCl, 60 mM MgCl2, 10 mM CaCl2, pH 7.9.
Enzyme Dilution Buffer: 25 mM Tris-HCl, 50% glycerol (v/v), pH 7.6 (at 4 °C).

Quality

Absence of contaminants: Each lot is tested to ensure the absence of RNases and proteases according to the current Quality Control procedures.

Specificity

Heat inactivation: One unit DNase I recombinant, RNase-free is heat-inactivated by 10 minutes incubation at 75 °C.

Important Note: Alternatively, DNase I recombinant, RNase-free can be inactivated and removed by phenol extraction according to standard protocols, e.g., Current Protocols in Molecular Biology.

Unit Definition

One unit is the enzyme activity that effects an absorbance increase of 0.001/minute under assay conditions in 1 ml at 260 nm.
Assay conditions:
Volume activity is determined according to the following assay mixture. 100 μg calf thymus DNA is incubated in 2.5 ml 1x incubation buffer with 40 to 70 units DNase I recombinant, RNase-free at +25 °C. The absorbance increase is measured at 260 nm.

Application

DNase I recombinant, RNase-free may be used to degrade DNA in applications that are sensitive to the presence of RNase. For example, DNase I is frequently used to:
• Remove genomic DNA from RNA preparations prior to RT-PCR
• Isolate DNA-free RNA after in vitro transcription reactions
• Perform nick translations
• Map DNase-sensitive regions in eukaryotic DNA

Specifications

Glycosylated form
Recombinant DNase I is heterogeneously N-glycosylated, so it appears as two bands in gel electrophoresis.
Divalent ion requirement
DNase I requires divalent cations for maximum activity. The DNA-specific endonuclease is activated by ions such as magnesium ions and is stimulated by calcium ions. Therefore, the enzyme is inhibited by metal chelating agents like EDTA.

Kit component only

Description

Product #

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Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
Protocols & Articles

Articles

3D Organoid Culture: New In Vitro Models of Development and Disease

Model systems drive biological research by recapitulating body processes and functions from the molecular to whole organism level. The human body is composed of both cellular and non-cellular materia...
Keywords: Adhesion, Cell attachment, Cell culture, Cryopreservation, Diseases, Growth factors, Infectious Diseases, Processes and Functions, Vitamins

Protocols

DNase I Recombinant, RNase-free Protocol

DNase I from bovine pancreas is a glycoprotein of Mr 37000. A special procedure is used to remove RNases from the DNase preparation. DNase I is a DNA-specific endonuclease that hydrolyzes ds or ssDNA...
Keywords: Centrifugation, Polymerase chain reaction, Precipitation

Peer-Reviewed Papers
15

References

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