Removal of template DNA following in vitro transcription (1).
Preparation of DNA-free RNA prior to RT-PCR and RT-qPCR (2).
DNA labeling by nick-translation in conjunction with DNA Polymerase I (1).
Studies of DNA-protein interactions by DNase I, RNase-free footprinting (1).
Generation of a library of randomly overlapping DNA inserts. Reaction buffer containing Mn2+ is used (3).
Description
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, see Fig.1:
– in the presence of Mg2+, DNase I cleaves each strand of dsDNA independently, in a statistically random fashion (1);
– 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 (1).
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