DNase I Powder

BR Grade, For Research Use Only

Cat. No. N9066

Size: 1 g

Specific Activity: ≥ 2000 U/mg

Form: Contains 1 g DNase I, lyophilized powder

Store at -20°C for 2 years.

Description

DNase I is an endonuclease that nonspecifically cleaves DNA to release di-, tri- and oligonucleotide products with 5´-phosphorylated and 3´-hydroxylated ends. DNase I acts on single- and double-stranded DNA, chromatin and RNA: DNA hybrids.

Applications

• Degradation of DNA template in transcription reactions

• DNase I footprinting

• Nick translation

Definition of Activity Unit

One unit of the enzyme completely degrades 1 µg of plasmid DNA in 10 min at 37°C. Enzyme activity is assayed in the following mixture: 40 mM Tris-HCl (pH 8.0), 10 mM MgSO₄, 1 mM CaCl₂, 1 µg of pUC19 DNA.

Source

Bovine pancreas.

Molecular Weight

38 kDa monomer.

Quality Control

Functionally tested for digestion of template DNA after in vitro transcription.

Inhibition and Inactivation

Inhibitors: metal chelators, transition metals (e.g., Zn) in millimolar concentrations, SDS (even at concentrations less than 0.1%), reducing agents (DTT and beta-mercaptoethanol), ionic strength above 50-100 mM.

Inactivated by heating at 65°C for 10 min in the presence of EGTA or EDTA (use at least 1 mol of EGTA/EDTA per 1 mol of Mn²⁺/Mg²⁺).

Note

• DNase I is sensitive to physical denaturation. Mix gently by inverting the tube. Do not vortex.

• This grade of DNase I is not used for removing DNA from RNA preparations.

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