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--
--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....-