Thermo Scientific 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 following in vitro transcription
• 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.