Application

Deoxyribonuclease I bovine has been used in a study to compare the initial actions of spleen deoxyribonuclease and pancreatic deoxyribonuclease. Deoxyribonuclease I bovine has also been used in a study to investigate deoxythymidine 3′, 5′-di-p-nitrophenyl phosphate as a synthetic substrate for bovine pancreatic deoxyribonuclease.

DNAse I from Sigma was used to treat nuclear lysate to obtain single nucleosomes in a study. The enzyme has also been for the preparation and harvest of mice mammary glands.

Used for the removal of DNA from protein samples.

Biochem/physiol Actions

DNase I is an endonuclease that acts on phosphodiester bonds (adjacent to pyrimidines) to produce polynucleotides with terminal 5′-phosphates. The pH optimum is found to be between 7 and 8. Divalent cations such as Mn²⁺, Ca²⁺, Co²⁺, and Zn²⁺ are activators of the enzyme. A concentration of 5 mM Ca²⁺ stabilizes the enzyme against proteolytic digestion. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin are known to inhibit the enzyme activity.

Digests single- and double-stranded DNA to a mixture of mono- and oligonucleotides carrying 5′ phosphates and 3′ OH termini. This catalytic activity is divalent ion-dependent. In the presence of Mg²⁺, DNase I hydrolyzes each strand of double-stranded DNA randomly and independently. In the presence of Mn²⁺, both strands can be cleaved.

Features and Benefits

RNA purification by removing DNA Prepare DNA for nick translation¹ Footprinting assays to determine DNA-protein interactions²

Physical form

Supplied as a solution in 4 mg/ml glycine pH 5.0, 5 mM calcium acetate and 50% glycerol

Preparation Note

Produced without using any animal cells or animal derived materials.

Unit Definition

One unit will produce a δA₂₆₀ of 0.001 per min per mL reaction mixture using calf thymus DNA at pH 5.0 and 25°C