Reversing crosslinks - (Jun/25/2010 )
By reading some of the forums in the archives, I stumbled under a post saying that it was possible to reverse crosslinks in DNA "by incubating the crosslinked solution in high salt (approximately 1ul 5M NaCl/25ul crosslinked material-upstate ChIP kit) at 65C for at least 4h" (Quote from beccaf22).
Is that true? Is it possible to reverse crosslinks without harming the DNA?
The only method I know of to break crosslinks is antigen retrieval but the heat kills the DNA .
What action would the NaCl have on DNA?
I guess that is pretty standard reverse crosslinking conditions. 4hr at 65C is a minimum, the incubation time could be longer.
OMG, and I have been looking for a solution for...6 years
Now I need to know more. Let's say you have a tissue that's been fixed a while ago, how do you proceed? You soak it in what concentration of NaCl? and then you rinse, and extract?? How does this work?
Do you know a good article I can use???