Re: [DIYbio] Re: Small side project - baking GFP bread - exhibtion

Just remember that some GFPs are sensitive to pH and will denature in acidic conditions. Maybe even the high temp of baking? Either way it would be neat to see if you can get decent expression. Best of luck!

Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D

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From: Mega [Andreas Stuermer]
Sent: 12/9/2013 2:11 PM
To: diy...@googlegroups.com
Subject: [DIYbio] Re: Small side project - baking GFP bread - exhibtion

As the fluorescent bread is approaching (hope to get it done before Christmas), I am thinking how I will breed the bugs.

I will innoculate about one liter of yeast medium from a petri dish.
In our lab exercise, we had professional fermenters. At home it would be possible to build one, as I have two air pumps lying around in the basement - just attach a sterile filter.

But I think the cheapest and quickest idea is to keep them in very big containers, with the liquid being just around two centimeters high. Thus free diffusion would be sufficient.

But regulating the pH value without contaminating it will be another issue. Wouldn't it be better to skip pH regulations? Add some Penicillin-G (it should decay during baking) and 8% ethanol. The ethanol will kill most of the bacteria anyway.

The initial pH would be around 5.5, so the yeast has an initial propagation advantage over many bacteria. The ethanol will do the rest.
After some time, the ethanol will be consumed aerobically by the yeast. But it will outgrow any bacteria before they get a chance to propagate I assume.

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