Blog Post Week 6

Figure 1. Beaker Method

Figure 2. Unsuccessful
Turkey Baster

We are still trying to find new a good protocol to follow for the microplastics. Since we could not get a funnel or any separation apparatus to work Matt suggested we tried the beaker method once more. This time the beaker method would be different in a way. Matt wanted me to get a known amount of plastic and mix it with our soil sample to see what percentage of the microplastics we could get back. I started with filing down plastic 

(I used a PVC Pipe as the plastic and a file to file the pipe) once I filed down enough plastic I weighed 1gram of it and 50 grams of soil and combined them together. I continued to add 200mL of NaCl solution to the sample and continue with the beaker method (figure 1). With the beaker method it states we transfer using a turkey baster but all the microplastics would get stuck on the side of the turkey baster making it unsuccessful (figure 2). I attempted to transfer the micropipette using a sterile transfer pipette but the microplastics clumped on the tip not allowing a proper transfer (figure 3). I tried with a micropipette and a regular green pump pipette and it did not work. Matt suggested that I pour the microplastics and the liquid into the other beaker (figure 4), He thought this would be the best way to transfer even though in was very inefficient. To completely transfer I had to add another 200 mL of solution to get the most microplastics, I added another 200mL and repeated my steps (total volume of 600mL of 5.0M NaCl Solution). Once I was done transferring the microplastics, I had to move on too filtering. I had concerns that our sample would not filter because of how dirty the sample was. Filtering did not go well either since the hose collapsed (figure 5). Everything that we have tried so far has not worked, we are going to keep trying. 

Pseudomonas update: We recently did gel electrophoresis and moved on to DNA extraction from the gel (figure 6).    

Figure 3. Clogged
Transfer Pipette 

                                                                                              

Figure 4. Transfer through pouring

Figure 5. Filtration/
 Collapsed hose

Figure 6. DNA extraction from gel