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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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chIP problem
Posted by: mb80 (IP Hidden, Junior member, 12)
Date: January 30, 2008 11:03PM
Hi,
I am having trouble with my chIP protocol. It worked twice in December, when I was able to get fold enrichments for my protein of interest of roughly 5.0 relative to IgG, and for Pol2 of 5.0-20.0, at my promoter of interest. In Jan, those numbers went down to <2.0 If anyone can give me input on how my sonications look, I would very much appreciate it--- "A" and "B" worked, "C" and "D" did NOT work well Do you think the relatively subtle differences between A/B and C/D could account for my troubles? Thanks for input!
Re: chIP problem
Posted by: drsgarrett (IP Hidden, New member, 3)
Date: April 4, 2008 03:36PM
Are A and B the December ones that worked?
If it worked one month and then didn't the next month, my best bet is that some of your reagents went off. If you're reasonably certain the antibodies are fine, I'd remake your buffers (assuming you aren't using a kit) and see if that works with a known system before going back to your experiment of interest. I doubt the sonication is an issue unless your sonicator broke.
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