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Team:Paris Bettencourt/Experiments/Methodologies
From 2011.igem.org
Methodologies
Creating nanotubes under the microscope need a lot of tuning. In these pages, we recapitulates the methodologies we use to prepare the microscope slides with the cell properly prepared, and how we have built the microfluidic chips in wich we trapped B.subtilis strains.
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Preparing the microscope slides: Visit this page if you want to learn how to grow B. subtilis strains in condition you can observe the nanotube.
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See our new microfluidic chip: We have modified a microfluidic system from Jeff Hasty group in UCSD, in which we can grow bacterial cells continuously for a long time in a single layer. We called it the Micro-chemostat. Compared to the usual microcolony-on-agar-pad method in the paper, this system will be able to maintain exponential growth of the bacterial cells in close contact with each other for a long time without exhausting the nutrient nor form double layers. We hope by extending the imaging time, we'll be able to observe the more diffusion-through-nanotube events in a single experiment.
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Integration in B.subtilis Integration of our constructs in B.subtilis proved to be a lot trickier than we expected. This might be the main reason for B.subtilis's lack of popularity in iGEM. You can find more about our struggles and how we solved this problem here. |
