DNase I recombinant, RNase-free

Description

• 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.ContentsRecombinant DNase I, RNase-free, 10 U/μl Incubation Buffer, 10x concentrated
• 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.
• Application: DNase I recombinant, RNase-free may be used to degrade DNA in applications that are sensitive to the presence of RNase.[1][2][3] For example, DNase I is frequently used to:Remove genomic DNA from RNA preparations prior to RT-PCRIsolate DNA-free RNA after in vitro transcription reactionsPerform nick translationsMap DNase-sensitive regions in eukaryotic DNA
• Features and Benefits: Eliminates DNA contamination from any RNA sampleContains no detectable RNase or protease activityCan be heat inactivated, thereby eliminating the need for organic extractionShipped with an optimized incubation buffer, which supports maximum DNase activityProduced via an entirely animal-free process, to eliminate any risks associated with animal-derived material
• Packaging: 1 kit containing 2 components
• 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 formRecombinant DNase I is heterogeneously N-glycosylated, so it appears as two bands in gel electrophoresis.Divalent ion requirementDNase 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.
• 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.
• Preparation Note: Activator: Bivalent metal ionsWorking 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.
• Other Notes: For life science research only. Not for use in diagnostic procedures.