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.
- Recombinant DNase I, RNase-free, 10 U/μl
- Incubation Buffer, 10x concentrated
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.
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
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
1 kit containing 2 components
Absence of contaminants: Each lot is tested to ensure the absence of RNases and proteases according to the current Quality Control procedures.
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.
One unit is the enzyme activity that effects an absorbance increase of 0.001/minute under assay conditions in 1 ml at 260 nm.
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.
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).
Storage and Stability
Store undiluted enzyme solution at -15 to -25°C; storage buffer at 4 °C.
For life science research only. Not for use in diagnostic procedures.