Dear all, I have had problems using benzonase nuclease to remove DNA from bacterial (E. coli) lysates, from which I am purifying a ssDNA-binding protein. The problem is, that I want to purify a DNA-free protein, and benzonase did work for that, but then of course I need to remove the benzonase from the preparation, because I later want to use this protein in experiments containing DNA, and benzonase proved to be difficult to completely remove. I still had nuclease activity in my "purified" protein. I am planning to try the cyanase nuclease from Ribosolutions Inc., since they claim it is faster than benzonase or DNase, and is more easily removed after use, with the "inactivation resin" that they provide with the enzyme. Does anyone have an opinion on what is your favorite nuclease for removing DNA from bacterial lysates, and that can then be removed itself also? DNase, benzonase, or cyanase..?
Thank you in advance, Paul Phelan Tufts University School of Medicine Biochemistry Boston _______________________________________________ Methods mailing list Methods@net.bio.net http://www.bio.net/biomail/listinfo/methods
