On the week of 10/6 we planned out the rest of our experiment that way we were prepared for the process. We figured out the salt concentration and decided on a serial dilution starting with 1M, to 0.1M, 0.01M, 0.001M concentrations. These concentrations are how we will be able to determine the effects of biofilm. The serial dilution started with an eppendorf tube filled with 1000 micro liters of stock salt. The picture below explains the concentrations in each.


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9/14 and 9/15 labs

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On 9/14 we did a gram stain on D. Rad. Unfortunately there were not enough cells and we did not heat fix long enough to see any result under the microscope. The cells began to wash away during the process. On 9/15 we decided to reattempt gram staining on P. fluorescens. This time we heat fixed even more so the cells did not wash away. The sample was a bit older coming from August, therefore the cells appeared as debris rather than clumps of cells.
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After about 3-4 weeks, we have ran 3 trials for each salt concentration. We have one set of data points so far and by Tuesday we will have all 3 trials figured out. The first round of data points all turned out good with no contamination. The process has been good overall and we have not ran into any major issues. In the beginning we had a few hiccups with contamination on our TGY plates but nothing since. We are excited to get the rest of our data points and run the statistics to put a poster together.
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On 9/28 we took 3 mL of TGY broth and placed it in a 15 mL flacon tube to prepare for inoculation on Saturday. Chad did a 4 way streak to begin growing the Gobi and allow to inoculate over night. On 9/29 we heated up the TGY broth that has agar to begin plating. The plates had some growth visible but no individual colonies. We did a gram stain on the culture that grow to look for grape like clusters of cells. Rebecca took a cluster of cells straight from the plates into a slide that has 5 micro-liters of TGY broth. The gram stain showed gobi was present but the heat fix caused the cells to turn black instead of purple.
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