User:Eric M. Walters/Notebook/Spring 2012/2012/05/04
|Cyanobacterial acrylate production||Main project page|
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Malonyl CoA reductase, why you gotta play me like that?
So it seems likely that the MCR is causing some sort of increased acrylate toxicity, likely due to the increase of CoA species in the cell. Before trying to transform again, I'm going to try to find the acrylate MIC in MCR-bearing 7002.
Today, I'm making plates of 5, 10, and 20 μM AA + Kan100 to grow them up on.
Gonna make 1 500 mL batch of kan+acrylate plates for growing up the transformants, once that happens
Plating the culture
The wt MCR culture was .348 OD. Given what Erin told me about plating efficiency (1 ODmL is 1e7 colonies), this should be confluent (3.5e5) below MIC. Putting 100 μL of culture onto each of 6 plates, duplicates of the 5, 10, and 20 μM acrylate with kan100.
Media A recipe
For 1 L of 1x cyanobacteria growth media
FOR THE 2x CONCENTRATION FOR PLATES: Don't add B12 (will be added later), and all above values are doubled.