Blog Post Week 8

For microplastics this week I tried the beaker method separation (figure 1) since I finally had shaved down enough plastic to attempt it. I was supposed to filter my sample once I was finished separating to see what percent of microplastic we could get back but the microbiology lab needed all the filtering equipment for the class. So I decided to leave the sample sitting and filter another day. When I came back to the lab to filter Matt had made a discovery! The sample had sat for two days. Our though was that all the plastics would float because they had a lower density than the 5M NaCl solution, but this did not happen (figure 2). Instead some of the plastics sank to the bottom. We know that the filed plastic is a microplastic so we had to find a solution with a higher density. This means that we have to find a cost effective solution. Matt suggested we attempt to make a 5M NaCl and 10% sucrose solution. This still did not have a high enough density, the highest density that we had was 1.301 but it was with a 5M NaCl and 60% sucrose solution (figure 3), which is not the best. We are once again stuck until we can find a good funnel to allow to do the separation and a solution with a greater density.

Figure 1. Beaker method Seperation 

Figure 2. Sample that was left out
Microplastic settled 

Figure 3. Attempt to make solution
with greater density

Pseudomonas Update: The microbiology labs did DNA extraction of their samples and we were in charge of nanodropping. The microbiology classes also ran a PCR and we were in charge of nanodropping that as well and screening them through gel electrophoresis.

Blog Post Week 7

This was a slow week in lab. I am behind on my background and method so I spent my time trying to catch up. I have also spent time filing down plastic. It takes me a long time to file down one gram of plastic to attempt the beaker method. Matt would like me to try the beaker method at the least five times. I have only down it once since I have been trying to file down as much plastic as possible.

Pseudomonas Update: The microbiology classes have started collecting water and biofilms sample. Our team is now in charge of using the class samples and taking the date. This week we took pictures of the plates and transferred some sample onto broth. We also began to organize a file that will include the classes data.

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