I have been working with yeast-2-hybrid for a while now and have been getting a lot of background growth, so much so that it is hard to detect genuine interactions. I have to use 3-AT and optimize its concentration. However I would like to have some suggestions on the best way to add 3-AT to my media. Should I add the powder directly to the media and autoclave it? Or should I make a stock solution and add it after autoclaving the media? Also any suggestions on how much 3-AT can be used to get rid of the background growth will also be helpful. Thanks!

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