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chIP assay


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#1 ulujm

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Posted 29 July 2009 - 05:49 AM

I was wondering why to crosslink?

by crosslinking every protein will be bind to DNA. so what the point since we want only the protein of interest.
by crosslinking you force the protein to bind even they don't bind. So you end up with protein binding to your promoter but is it a real binding or the result of cross linking.

am i right?

#2 Dr Teeth

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Posted 29 July 2009 - 06:07 AM

I was wondering why to crosslink?

by crosslinking every protein will be bind to DNA. so what the point since we want only the protein of interest.
by crosslinking you force the protein to bind even they don't bind. So you end up with protein binding to your promoter but is it a real binding or the result of cross linking.

am i right?


Crosslinking is not total. Proteins in the vicinity of DNA will crosslink to the DNA, which would not be the case for most proteins. Also, specificity is achieved through repeated experimentation. A protein that just "happens" to be near your promoter region, but is not specific, will not likely be seen in repeated experiments. Also, as is the case with all experiments, results should be supported by other data as well to become meaningful.

Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley

#3 ulujm

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Posted 29 July 2009 - 06:12 AM

so what about the reverse crosslink? it should unbind everything right?

or only a strong binding protein will stay?

#4 cellculturenovis

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Posted 27 July 2010 - 06:36 AM

You should only reverse crosslink an aliquot of your sample in order to quantify the DNA. You can then proceed with the cross linked sample to the IP stage.




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