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no DNA after IP


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#1 Travis

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Posted 24 July 2009 - 07:46 AM

Hi, I am having problems with ChIP. I really really need help fomr you guys. I have posted several months before about how i have too contaminated ChIP samples after IP. Well then I discover that theres actually no DNA in my sample at all. So simply speaking, after IP, I have no DNA. I am using Protein A/G agarose beads, and mouse monoclonal Ab (not ChIP grade). I am using a fast ChIP protocol and I prepare IP buffers myself. I have no luck in obtaining DNA. Why is it like this? Has anybody experience this?

#2 KPDE

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Posted 24 July 2009 - 12:55 PM

Hi, I am having problems with ChIP. I really really need help fomr you guys. I have posted several months before about how i have too contaminated ChIP samples after IP. Well then I discover that theres actually no DNA in my sample at all. So simply speaking, after IP, I have no DNA. I am using Protein A/G agarose beads, and mouse monoclonal Ab (not ChIP grade). I am using a fast ChIP protocol and I prepare IP buffers myself. I have no luck in obtaining DNA. Why is it like this? Has anybody experience this?



Are you using a positive control antibody along with positive control primers (i.e. an antibody that you know works well in ChIP and a primer pair where you know the protein of interest binds)? If not, this is where you should start.




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