Progress (week 5)

Figure 1a: PVC 3 plastic grinding with partner Sky

            Progress is being made on our WAESO research regarding microplastics. This week, we finally started grinding our different types of plastics. We decided to start with our plastic pipe (PVC 3) because it would be easier to grind due to its rigid and flexible properties (figure 1a). The grinding process will take a couple of weeks, which is good because we will have more time on our bacteria’s. Our different types of Pseudomonas bacteria grew well. Now it’s time for DNA extraction. DNA extraction protocol, in short, is for separating the desired plasmids from its components, which are RNA, protein, chromosomal DNA, etc.

Buffer solutions for DNA extractions

Before extracting, four types of solutions/buffers should be prepared (figure 1b). The first solution prepared was “100% Isopropanol.” Since DNA is insoluble in ethanol and “100% isopropanol”, the addition of alcohol, followed by centrifugation, will cause the DNA proteins to come out of the solution. The second solution prepared was 7.5 M of ammonium acetate. Ammonium acetate is used to remove cellular proteins bounded to the DNA. The third buffer made was 70% ethanol. Ethanol is a basic technique used for concentrating and de-salting nucleic acids (DNA or RNA) preparations in aqueous solution, which forces the precipitation of nucleic acids out of the solutions. The fourth and final buffer made was the Lysis buffer. A Lysis buffer (obvious from its name ‘lysis’) is a buffer solution used for breaking open cells. These four solutions/buffers are ready for use once we get to the lab tomorrow (9/29/2017) for DNA extraction.