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D2821

Sigma-Aldrich

Deoxyribonuclease I bovine

recombinant, expressed in Pichia pastoris, lyophilized powder, RNAse and protease, free

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Synonym(s):
DNAse I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
Enzyme Commission number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54

biological source

bovine

Quality Level

recombinant

expressed in Pichia pastoris

form

lyophilized powder

specific activity

≥4,000 units/mg protein

mol wt

~39 kDa

technique(s)

DNA extraction: suitable

solubility

H2O: soluble (pH 4.0-9.0)

suitability

suitable for molecular biology

application(s)

diagnostic assay manufacturing

foreign activity

RNAse and protease, free

storage temp.

2-8°C

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Application

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.
Deoxyribonuclease I bovine has been used in the preparation of cold cell lysis buffer, complete RNA lysis buffer, cell lysis buffer for testing the expression of recombinant tagged protein.
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 expression of KNOTTED1-like HOMEOBOX (KNOX) transcription factors.
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 lyophilized powder containing glycine as a stabilizer

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.

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

常规特殊物品

Certificates of Analysis (COA)

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Adele Williamson et al.
Protein expression and purification, 97, 29-36 (2014-03-04)
The genome of the psychrophilic fish-pathogen Aliivibrio salmonicida encodes a putative ATP-dependent DNA ligase in addition to a housekeeping NAD-dependent enzyme. In order to study the structure and activity of the ATP dependent ligase in vitro we have undertaken its
Recombinant expression and purification of an ATP-dependent DNA ligase from Aliivibrio salmonicida.
Williamson A and Pedersen H
Protein Expression and Purification, 97(4), 29-36 (2014)
Quantitative proteomics of breast tumors: Tissue quality assessment to clinical biomarkers.
Chen Y, et al.
Proteomics, 17(6), 1600335-1600335 (2017)
Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening.
Joung J, et al.
Nature Protocols, 12(4), 828-828 (2017)
Properties of chromatographically purified bovine pancreatic deoxyribonuclease.
P A Price et al.
The Journal of biological chemistry, 244(3), 917-923 (1969-02-10)

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