Deoxyribonuclease I Product Information

I.U.B.: 3.1.21.1

Bovine pancreatic deoxyribonuclease is an endonuclease which splits phosphodiester linkages, preferentially adjacent to a pyrimidine nucleotide yielding polynucleotides with free hydroxyl group at the 3' position and phosphate group at the 5' position. The average chain length of a limit digest is a tetranucleotide.

Uses: Worthington offers DNase at different levels of purity for different applications. Product Codes: DPRF and DPRFS are both especially designed for Molecular Biology applications and contain the lowest levels of RNase and protease activity. They are both suitable for use in techniques requiring digestion of DNA in the recovery of intact RNA or where the integrity of structural proteins or enzymes must be maintained. Applications have included nick translation, DNA mapping, isolation of nuclear RNA and protein, plasmid construction, and RNA polymerase synthesis of RNA probes and RT-PCR. DNase is also used in tissue culture work to digest DNA from damaged cells thereby reducing viscosity, and removing membrane bound DNA fragments. Worthington Codes: DP and DCLS are suitable for these applications.

Stability/Storage: When properly stored, all grades of Worthington deoxyribonuclease are stable for 2-3 years. Recommended storage temperature for all grades of Worthington DNases is 2-8°C. Product code DPRFS may be stored at -20°C. For long term storage in solution, Product Codes D and DPFF may be dissolved in 5mM acetate, 1mM calcium, pH 4.5 and stored in single use aliquots at -20°C or -70°C for up to one year. Only freeze and thaw once; thawed aliquots are stable refrigerated at least several weeks. Addition of 50% glycerol will maintain a liquid state at -20°C without affecting stability and material in 50% glycerol can be removed and returned to -20°C repeatedly. DPRF is unusually stable due to the absence of protease. For long term storage of DPRF after reconstitution, use water or any buffer pH 4.0 to 9.0 except phosphate; avoid calcium chelators; add 50% glycerol for storage as liquid at -20°C; aliquot in single use containers; only freeze and thaw once; thawed aliquots are stable refrigerated at least several weeks.

Unit Definition: 1 Unit causes an increase in absorbance at 260nm of 0.001 per minute per ml at 25°C when acting upon highly polymerized DNA at pH 5.0. Note: Kunitz units as reported by other suppliers can be 2 to 4 times higher than Kunitz units as measured at Worthington. As measured at Worthington, one Kunitz unit digests 1µg of lambda DNA in 10 minutes at 37°C in 50mM Tris, 1mM Mg²⁺, 1mM Ca²⁺, pH 7.8 in a 50µl reaction. Correlation of digestion units with Kunitz units is different for other DNA and buffer systems.

Technical Note: Product Code DPRF: Each vial contains approximately 2mg glycine and 2µmoles calcium per 10,000 units of DNase I. Dissolving the entire vial in 5ml provides the equivalent of a 1 mg/ml solution.

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