Merck, D5319-2MG, Deoxyribonuclease I bovine

Application
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.
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.
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 acetateand 50% glycerol
Preparation Note
Produced without using any animal cells or animal derived materials.
Other Notes
One unit will produce a ΔA₂₆₀of 0.001 per min per mL reaction mixture using calf thymus DNA at pH 5.0 and 25°C