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 (see Reference 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 (see Reference 1).
- Recombinant enzyme
- Purified from non-animal host with a lower level of intrinsic RNases
- Preparation of DNA-free RNA (see Reference 1)
- Removal of template DNA following in vitro transcription (see Reference 1)
- Preparation of DNA-free RNA prior to RT-PCR and RT-qPCR (see Reference 2)
- DNA labeling by nick-translation in conjunction with DNA Polymerase I (see Reference 1)
- Studies of DNA-protein interactions by DNase I, RNase-free footprinting (see Reference 1)
- Generation of a library of randomly overlapping DNA inserts. Reaction buffer containing Mn2+ is used (see Reference 3)
DNase I is sensitive to physical denaturation. Mix gently by inverting the tube. Do not vortex.
|10X Reaction Buffer with MgCl2||100 mM Tris-HCl (pH 7.5 at 25°C), 25 mM MgCl2, 1 mM CaCl2.|
|10X Reaction Buffer without MnCl2||100 mM Tris-HCl (pH 7.5 at 25°C), 1 mM CaCl2. Recommended concentration of MnCl2 in 1X reaction buffer is 10 mM.|
|Definition of Activity Unit||
|Inactivation||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+ (5)).|
|Inhibition||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 to 100 mM.|
|Molecular Weight||29 kDa monomer|
|Source||E. coli cells with a cloned gene encoding bovine DNase I.|
|Storage Buffer||The enzyme is supplied in:
50 mM Tris-HCl (pH 7.5), 10 mM CaCl2, and 50% (v/v) glycerol.
|Storage Condition||-20 C|
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