2014hs.igem.org/team:Charlottesville RS/Notebook/041814

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April 18th, 2014

Alli Ambrosini

Did research on how to test the results of our project. The team will need to get non-nutrient agar plates so that it can be tested whether the bacteria can eat our PHB at all. Normal agar has dextrose in it, which bacteria will consume before they touch PHB because its easier to eat. A simple sugar is a lot easier to break down than a long polymer such as PHB.

Lauren Ewell, John Grammer

Took competent cells, centrifuged them to get a pellet. Resuspended in Luria broth, centrifuged and then combined into three tubes and added calcium chloride and put on ice for 20 minutes. Alli centrifuged gently by pulsing 10 times and then resuspended with 200uL of cold calcium chloride.

Alex Manchester, Nick Keen, Anders Beaurline, Jessica Prax, Bailey Fernandez

Did the re-suspending of DNA and transformation procedure.

The cells were possibly not competent because they were not suspended, there was a pellet at the bottom

The transformation procedure in the iGEM procedure notebook on page 28 was performed There were not 2 hours to incubate it, which was step 6

Mrs. Minutella plated the transformed DNA at 60uL on each plate

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