Are DNase inhibitors available? - help (Apr/06/2006 )
I am trying to identify DNA binding proteins from Streptomyces.I am using Biotinylated DNA bound to Streptavidin magnetic beads for this purpose.When i add crude cell lysate to beads I find degradation of the bound DNA.I have used 1mM EDTA in the buffer which i use for binding.
Can anybody help me out with this ..?and let me know if DNase inhibitors are commercially available.
Thanks in advance.
i don't know any inhibitor, but if you would follow a protocol for EMSA you should not have problems with DNase.
You have to isolate nuclear proteins and do not use the total extract, I would say.
I agree; isolating a nuclear protein extact would almost certainly reduce the amount of DNase activity. Keep everything on ice as well.
Are you positive it's DNase activity and not some other part of you protocol which might cause the degradation?
Some info here...