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How do you reverse formaldehyde crosslinking? - (Oct/19/2005 )

Hello,
I am haivng difficulities finding a protocol for the reversal of formaldehyde crosslinking. I am examining interaction between crude protein and a specific sequence of DNA, I would like to crosslink them, however I also need to separate them. If anyone knows of a "reversible" protocol plase let me know.
Thank you

-mpage-

formaldehyde crosslinking is reversed by incubating the crosslinked solution in high salt (approximately 1ul 5M NaCl/25ul crosslinked material-upstate ChIP kit) at 65C for at least 4h

--someone else should confirm this, but I think this reverses imino bonds formed between the transcription factor and the DNA by the formaldehyde--

-beccaf22-

Yes beccaf22 is right. Carry out the reverse crosslinking AT LEAST for 4hr.

-pcrman-

QUOTE (beccaf22 @ Oct 19 2005, 09:53 PM)
formaldehyde crosslinking is reversed by incubating the crosslinked solution in high salt (approximately 1ul 5M NaCl/25ul crosslinked material-upstate ChIP kit) at 65C for at least 4h

--someone else should confirm this, but I think this reverses imino bonds formed between the transcription factor and the DNA by the formaldehyde--



I leave to 65 ºC overnight.

-Research will kill me....-