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D5319

Sigma-Aldrich

Deoxyribonuclease I bovine

recombinant, expressed in Pichia pastoris, buffered aqueous glycerol solution, ≥5,000 units/mg protein

Synonym(s):
DNAse I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
Enzyme Commission number:
MDL number:
NACRES:
NA.54

recombinant

expressed in Pichia pastoris

Quality Level

form

buffered aqueous glycerol solution

specific activity

≥5,000 units/mg protein

mol wt

~39 kDa

foreign activity

RNAse and protease, free

shipped in

dry ice

storage temp.

−20°C

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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

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

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.

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

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Certificate of Origin

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