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Spatiotemporal redox signaling apparatus

NB Narendranath Bhokisham
EV Eric VanArsdale
KS Kristina T. Stephens
PH Pricila Hauk
GP Gregory F. Payne
WB William E. Bentley
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We used an Owl Easycast B1A (Thermo Scientific) gel electrophoresis system in the anaerobic chamber to create electrochemical gradients. We degassed the minimal-M9 media by constant stirring under anaerobic conditions and added 200 ml of degassed media into the electrophoresis gel system. Later, we supplemented the media with 50 mM potassium ferrocyanide and 5 µM PYO. To create a gradient, our electrochemical setup consisted of a working electrode connected to the left chamber of the device, a counter electrode connected to the right chamber of the device, and an Ag/AgCl reference electrode placed near the working electrode in the left chamber of the device. Reference was isolated from the media by a low pass frit. All the electrodes were connected to a CHI 620E potentiostat and a desired electric charge was applied.

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