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DNase I (RNase-Free), 1g

Price: $385.00

Specific Activity: ≥ 2000 Kunitz units/mg
 
Form: Lyophilized Powder
 
Storage: 4oC
 
Description
DNaseⅠ(RNase-free) 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.
 
Feature
  • Recombinant enzyme
  • Purified from non-animal host with a lower level of intrinsic RNases.
Applications
  • Degradation of DNA template in transcription reactions
  • Removal of contaminating genomic DNA from RNA samples
  • DNase I footprinting
  • Nick Translation
Quality Control
The absence of ribonucleases is confirmed by appropriate quality test. Functionally tested for digestion of template DNA after in vitro transcription
 
Source: A E. Coli strain that carries an MBP fusion clone of Bovine Pancreatic DNase I.
 
Molecular Weight: 29 kDa monomer
 
Definition of Activity Unit
One unit of the enzyme completely degrades 1 μg of plasmid DNA in 10 min at 37oC. 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.
 
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 65oC 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.
 
Preparation Instructions
DNase I is soluble in 0.15 M NaCl at 5 mg/ml. Solutions of DNase I at 5 mg/ml in 0.15 M NaCl may lose 10% of its activity when stored for a week in aliquots at –20oC. The same solutions stored in aliquots at 2–8oC can lose 20% activity.
 
Alternatively, solutions at a minimum concentration of 1 mg protein/ml, with 5 mM calcium chloride and stored at –20oC, may retain 90% activity for at least a year. Solutions containing 0.1 mg protein/ml are considerably less stable, and may require gelatin as stabilizer.
 
For applications unaffected by glycerol, two other storage buffers are options, as these formulations do not freeze at –20oC.
  1. 20 mM sodium acetate (pH 6.5), containing 5 mM CaCl2 and 0.1 mM phenylmethylsulfonyl fluoride (PMSF), 50% (v/v) glycerol, with DNase I at 5 mg/ml.
  2. 10 mM Tris-HCl (pH 7.5), 50 mM NaCl, 10 mM MgCl2, 1 mM DTT, 50% (v/v) glycerol, with DNase I at 2 mg/ml.

 


Add to Cart:

  • Model:
    [EPD01-02]
  • Shipping Weight: 20lbs

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