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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