Week 12 harbors the first bold steps into the heart of the experimentation; the investigation of the core hypothesis. The week began by creating large broth cultures of E.Coli to later be innoculated with the T2 and T4 phage cultures so that the phages may replicate within the E.Coli bacteria and we may retrieve a stock of phage solution for later use. The broths were created and set to grow in the incubator. Before the E.Coli broths were turbid enough to use to make the stock solutions, a small sample was taken and placed in two snap tubes, one was innoculated with T4 phage culture, and one with T2. To keep the solution moving and viable, a contraption was innovated which involves a stir bar and a small amount of water to agitate the snap tubes since we unfortunately didn't have a shake plate at our disposal. These will grow and be used as a preliminary test set to see how effective and efficient the phages are at ridding solutions of E.Coli contamination. The contents of the snap tubes were streaked out after many hours and the growth will be analyzed in the coming days. With the information gained from these experiments we will edit the rest of our procedure for the experimentation to fit our findings and streamline our testing.
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Week eleven has been a lot of preparation. While these steps are extremely important, it has been a lot of the same activities. We began by pouring agar into the amount of plates that will be carrying us through the first few rounds of experimentation (pictured below). This was accomplished by a day of mixing and cooking the agar recipe, autoclaving (sterilizing) it and pouring the actual plates out. Along with plates, we poured about 30 agar slants in which we will grow the initial E.Coli colonies. The tank continued to settle in and get re-oxigenated via bubbler (pictured), the established flora and mineral levels will continue to stay within normal pond levels as long as there is some form of oxygenation and current. We streaked the E.Coli onto the new slants, as we will be doing every 3-4 days to keep them fresh, and made sure the incubator was working; it was, and the E.Coli are prospering. Due to midterms, we had to work our schedule around the coming week so there will not be any active, or major, experimentation, and therefore no blog updates, until the week after next. You can stay up to date with a daily account of all laboratory activity by viewing the team's Daily Experimentation Log (Link on the homepage).
The team has been planning and coordinating with all the people involved with ordering the supplies for the past few weeks. It has been a few weeks of the same stuff, hence no updates (Sorry). After getting approval for using the grant money, finalizing the order with Mrs.Peterson, and actually ordering the supplies the waiting period began. Trying to stay productive during such a period, the team traveled to Sylvan Glenn beach to obtain some pond water to set up the testing environment. Using a large fish tank courtesy of Mr.Ellis, the pond water, and some soil/sediment from the pond, along with a bubbler to keep the water aerated, the environment was alive and ready (pictures below). Throughout this time a procedure also had to be made so day by day as we are moving through the experiment, we can know what the logical order of things is; however, the main idea is we use the pond water and periodically infect portions of it with Eschericia Coli, we then use bacteriophages to kill the E.Coli, varying the amounts of each to deduce the best starting concentration and conditions, and then devise a way in which this would be practically applied to a real-world body of water to cure an E.Coli outbreak more quickly and effectively. On Friday, January 6th 2017, the package arrived, it was just like a second Christmas morning! All the materials were unboxed and prepped for use in the coming weeks (Pictures below). In the package was petri dishes, sterile filtration units, a water test kit, "care and keeping of" handbooks, and of course, a vial of E.Coli K12 strain, a vial of Bacteriophage T2 strain and a vial of Bacteriophage T4 strain. Testing updates to come!
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