S-STEM Scholar Baylee Herman's blog

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.

The week of 10/11 and 10/12, Becca and I filled our 96 well plates with 10 micro liters of salt and 90 micro liters of our TGY with Gobi. After this was done, we had to let the bio film grow for 4-5 days. After the growth the next steps were doing a water wash, the CV, and the acetic acid. The process for the water wash and Crystal Violet is to sling, wash with water, pat dry, and repeat 3 times.

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.

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.