DNase I recombinant, RNase-free
ROCHE/04716728001 - from bovine pancreas, expressed in Pichia pastoris
Product Type: Chemical
DNase I recombinant, RNase-free
| biological source | bovine pancreas |
| form | solution |
| manufacturer/tradename | Roche |
| mol wt | ~39 kDa |
| optimum pH | 7.0-8.0 |
| packaging | pkg of 10,000 U |
| Quality Level | 100  |
| recombinant | expressed in Pichia pastoris |
| 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 |
| 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. |
| 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. |
| 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. |
| Storage and Stability: | Store undiluted enzyme solution at -15 to -25°C; storage buffer at 4 °C. |
| 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. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| Enzyme Commission (EC) Number | 3.1.21.1 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352200 |