So, it turns out that the protein I'm trying to purify is only soluble when I overexpress it in a one liter culture, but completely insoluble when I grow the cells in any larger volume. This makes no sense. Stupid protein.
You need to realize this is also an enzyme which (once I purify the darn thing) is only active anaerobically, at temperatures above 70°C, with 0.5M phosphate, in the dark.
Oh yes, I'm certainly looking forward to the next few months. Yes I am.
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