New England Biolabs, B0303S, DNase I Reaction Buffer

Related Categories Exonucleases and Non-specific Endonucleases,, Buffers Applications RNA Cloning Specification 1X Buffer Components 10 mM Tris-HCl 2.5 mM MgCl2 0.5 mM CaCl2 pH 7.6@25°C FAQ 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: Do you recommend a cleanup step after a DNaseI reaction? A: Yes. Depending on your downstream applications, we recommend a phenol/chloroform extraction step to remove all traces of the enzyme. The heat inactivation step is sufficient for inactivation of the enzyme, but will not entirely remove the enzyme. Q: Can I use an RNase inhibitor with my reaction? A: Yes. We recommend that you do to prevent any RNase from interfering with your reaction.