{"product_id":"neb-m0303l","title":"New England Biolabs, M0303L, DNase I (RNase-free)","description":"\u003cb\u003eRelated Categories\u003c\/b\u003e Chromatin Analysis,, Exonucleases and Non-specific Endonucleases,, RNA Synthesis In vitro Transcription (IVT),  \u003cb\u003eApplications\u003c\/b\u003e RNA Cloning  \u003cb\u003eSpecification\u003c\/b\u003e \u003cb\u003eMaterials Required but not Supplied\u003c\/b\u003e Nuclease-free Water (NEB #B1500) RNase Inhibitor, Murine (NEB #M0314) EDTA  \u003cb\u003eUnit Definition\u003c\/b\u003e One unit is defined as the amount of enzyme which will completely degrade 1 µg of pBR322 DNA in 10 minutes at 37°C in DNase I Reaction Buffer. Complete degradation is defined as the reduction of the majority of DNA fragments to tetranucleotides or smaller.  \u003cb\u003eReaction Conditions\u003c\/b\u003e 1X DNase I Reaction Buffer Incubate at 37°C 1X DNase I Reaction Buffer 10 mM Tris-HCl 2.5 mM MgCl2 0.5 mM CaCl2 (pH 7.6 @ 25°C)  \u003cb\u003eUsage Concentration\u003c\/b\u003e 2,000 units\/ml  \u003cb\u003eStorage Buffer\u003c\/b\u003e 10 mM Tris-HCl 2 mM CaCl2 50% Glycerol pH 7.6 @ 25°C  \u003cb\u003eHeat Inactivation\u003c\/b\u003e 75°C for 10 minutes  \u003cb\u003eFAQ\u003c\/b\u003e Q: What is the specific activity of DNase I(RNase-free)? A: The specific activity is approximately 20,000 Units\/mg. Q: Will DNase I work in NEB buffers 1-4? A: Yes, but not as efficiently. DNase I is a Calcium dependent enzyme so it works best in it’s own buffer which contains Calcium Chloride. It is recommended that even if you are using NEB buffers 1-4 that you also add DNase I buffer or .5mM CaCl2 to the reaction. Also, DNase I does not like high salt, as contained in NEB buffer 3. It works least efficiently in buffer 3, but will still give 50-75% digestion. Q: What is the best way to remove DNase I from my reaction? A: The best way to remove DNase I from your reaction is to perform a phenol\/chloroform extraction or to use a spin column. You can do the heat inactivation step, but that may not completely remove all of the DNase I, and it could interfere with your downstream applications. Q: Will DNase I work in rCutSmart buffer? A: Yes, DNase I will work in rCutSmart, with the addition of calcium. To see its % functional activity in rCutSmart, and that of other DNA Modifying enzymes in the cloning workflow, refer to the Activity of DNA Modifying Enzymes in rCutSmart® Buffer chart. Q: Can I use the Monarch Spin RNA Cleanup Kit to cleanup up my DNase I-treated RNA? A: Yes, the Monarch Spin RNA Cleanup Kits will remove residual DNase I (NEB #M0303) activity from RNA samples. On-column treatment with DNase I can often result in residual DNase I activity, therefore, we do not recommend on-column DNase I treatment with the Monarch Spin RNA Cleanup Kit, but rather in-solution DNase I treatment prior to RNA cleanup. ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835495862441,"sku":"M0303L","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-m0303l","provider":"Iright","version":"1.0","type":"link"}