Team:Imperial College London/Extras/Protocols/Auxin

From 2011.igem.org

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<h1>Auxin Lab Protocols</h1>
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<h2>27th of July</h2>
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The strain of E. coli with the copy of super-folded GFP that was incubated the previous night has shown limited/no growth in the prepared LB broth. This could have been caused by:<br>
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<h1>Auxin Xpress</h1>
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-No innoculation<br>
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<h2>Effect of Auxin on Plants</h2>
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-Kanamycin concentration in LB broth was too high. We had used 85μg/ml.<br><br>
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<b>New protocol:</b><br>
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<h2>Split Root Experiment</h2>
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50μg/ml is the recommended concentration. We will use 35-40μg/ml of kanamycin to speed up growth.</html>
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<h2>Salkowski</h2>
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<h2>28th of July</h2>
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Transformation of cells with 6, 7 and 8:<br><br>
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<h2>HPLC</h2>
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-Let competent cell strain 5α thaw for around 10 minutes on ice.<br>
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-Add 2-3μl of DNA.<br>
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<h2>Soil Erosion Experiment</h2>
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-Leave on ice for 20-25 minutes.<br>
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-Heat shock cells at 42°C for 45 seconds.<br>
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-Leave on ice for 10 minutes.<br>
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-Add 500μl of LB broth.<br>
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-Incubate for 1 hour at 37°C.<br>
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-Centrifuge for 1 minute.<br>
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-Remove 100μl off the top of the eppendorf tube. Pour out the rest making sure that the pellet remains in the eppendorf tube.<br>
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-Re-suspend the cells in the 100μl LB broth solution that was removed in the previous step.<br>
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-Add 5μl on a chloramphenicol agar plate (concentration of 35μg/ml).<br>
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-Add the rest of the sample to a second chloramphenicol agar plate.</html>
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Latest revision as of 14:25, 14 September 2011



Protocols

This page lists all the protocols used in our project. We have classified them into five main categories as follow.



Auxin Xpress

Effect of Auxin on Plants

Split Root Experiment

Salkowski

HPLC

Soil Erosion Experiment

Retrieved from "http://2011.igem.org/Team:Imperial_College_London/Extras/Protocols/Auxin"