Hi everyone! I'm performing a ChIP-Seq in tissues in a laboratory 4 hours away from mine so, I wanted to know if it's possible to keep the tissue frozen until the experiment is ready to be done in the other lab or if i have to perform the ChIP-Seq assay immediately after tissue sampling wich involves animal transportation.
Another thing that would be really helpful: if someone has permormed a ChIP-Seq in amphibian tissues, how much of it have you used to acquire the best results?
Thank you!
I second Milos that performing it in one go is optimal. Albeit not always possible. In your case I would suggest you perform tissue disruption, cross linking AND cell lysis in the place where you harvest the tissue. You will need minimal reagents for that. The resulting cross-linked nuclei can then be frozen and taken directly to the next step, which is chromatin shearing. I would advise against freezing whole cells. In my experience they become very fragile after fixing and freeze-thaw cycle leads to big material loss. I would also advise again freezing the whole tissue. This can lead to all sorts of problems.
As far as the amount of tissue to use, I would suggest to think about it in terms of cells. Disrupt the tissue to a single cell suspension (or near enough). Count the cells and proceed accordingly. Most ChIP protocols are based on about 10M cells. I would say a comfortable minimum is 1M.
+1 for Chipmentation. I really like the idea and it worked well in my hands as well.
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