I'm working on Methylation/Demethylation of certain promoter regions in DT-40 cell genome. Any recommendations as to folic acid/5-azac concentration and exposure time for that type of cell culture? I've ssen some articles using 4 mg/l and 40 mg/l but on a different cell line. Thoughts?
For the 5-azacytidine, 10 uM seems pretty standard. I recently purchased this product from Sigma, you may find it interesting: http://www.sigmaaldrich.com/catalog/product/sigma/a1287?lang=en®ion=US. As for the folic acid, I would check the concentration in your media, and then double this (there are also some low folate media available). You will need to treat for at least one cell doubling for a short treatment, 2-3 doubling for a solid response. 5-azacytidine is very unstable in solution, so you may want to freeze it in small aliquots and change your media at least every day.
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