Product Name

Deoxyribonuclease I bovine

Synonym

DNAse I, Deoxyribonucleate 5′-oligonucleotido-hydrolase

Assay

≥4,000 units/mg protein; recombinant, expressed in Pichia pastoris

Molecular Weight

mol wt ~39 kDa

MDL number

MFCD00130918

Suitability

BioReagent

l Physical and Chemical Information

Appearance

White lyophilized powder

Physical form

supplied as a lyophilized powder containing glycine as a stabilizer

Solubility(25°C)

H²O: soluble (pH 4.0-9.0)

l Biological Information

Biochem/Physiol Actions

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.

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.

Application

Used for the removal of DNA from protein samples.

Deoxyribonuclease I bovine has been used in a study to investigate the inhibition of DNA polymerase by extracts of rat liver. Deoxyribonuclease I bovine has also been used in a study to investigate the effects of ionic strength on enzymic activity.

The enzyme from Sigma has been used for the digestion of DNA extracted from Agave spp. clones. The enzyme was used during a study that investigated the effect of epigenetic changes on the regulatory  of KNOTTED1-like HOMEOBOX (KNOX) tranion factors.

Preparation Note

Produced without using any animal cells or animal derived materials. The enzyme powder may be reconstituted in water or any buffer at pH 4.0-9.0, except phosphate buffer. Calcium chelators should be avoided. 10 mg/mL solution of DNAse I in 0.15 M NaCl may lose <10% of its activity when stored for a week in aliquots at –20 °C. The same solutions stored in aliquots at 2-8 °C can lose approximately 20% activity. It remains active for upto five hours at 60 °C and loses activity in <10 minutes at 68 °C. It loses activity at the rate of 6%/hour in acetate buffer (pH 5.0) and tris buffer ((pH 7.2) at 1 mg/mL concentration.

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

l Storage

Storage temp.

2-8°C

l Precautions and Disclaimer

This product is for R&D use only, not for drug, household, or other uses.

l References

1. http://www.drugbank.ca

2. https://ncit.nci.nih.gov

3. https://www.ncbi.nlm.nih.gov