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Thermo Scientific™ DNase I, RNase-free


An endonuclease that nonspecifically cleaves single- and double-stranded DNA to release di-, tri-, and oligonucleotides with 5'-phosphate and 3'-OH groups.

Manufacturer: thermo scientific™  EN0525

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Catalog No. FEREN0525

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Description & Specifications

Specifications

Concentration 1U/µL
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
Source E.coli cells with a cloned gene encoding bovine DNase
Molecular Weight 29 kDa monomer
Quantity 1U/µL (Supplied with MnCl2)
Units 1,000U

Dnase I, Rnase-free is an endonuclease that digests single- and double-stranded DNA. It hydrolyzes phosphodiester bonds producing mono- and oligodeoxyribonucleotides with 5'-phosphate and 3'-OH groups.

The enzyme activity is strictly dependent on Ca2+ and is activated by Mg2+ or Mn2+ ions.

  • In the presence of Mg2+, Dnase I cleaves each strand of dsDNA independently in a statistically random fashion.
  • In the presence of Mn2+, the enzyme cleaves both DNA strands at approximately the same site, producing DNA fragments with blunt-ends or with overhang termini of only one or two nucleotides.

Highlights

  • Recombinant enzyme
  • Purified from non-animal host with a lower level of intrinsic RNases

Applications

  • Preparation of DNA-free RNA
  • Removal of template DNA followingin vitrotranscription
  • Preparation of DNA-free RNA prior to RT-PCR and RT-qPCR
  • DNA labeling by nick-translation in conjunction with DNA Polymerase I
  • Studies of DNA-protein interactions by DNase I, RNase-free footprinting
  • Generation of a library of randomly overlapping DNA inserts. Reaction buffer containing Mn2+ is used

Note

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