Starting PCR

After waiting a week for the primers to arrive, I was finally able to start PCR. However, before I began, I had to prepare my primers. By doing so, I had to centrifuge them, add the designated amount of Tris Hydrochloride to each primer (Figure 1) and shake like crazy! This process took about 4 hours because I had to make sure my primers were completely mixed with the Tris Hydrochloride (thus, ready to use). After I prepared my primers, I set up my samples for PCR. Each PCR tube required 10 microliters of 2X Master Mix for PCR, 2 microliters of the forward primer, 2 microliters of the reverse primer, 4 microliters of sterile water and 2 microliters of DNA (from my bacteria samples). For this PCR run, I used the PCAT-4f-2015 and PCAT-4r-2015 primers (they are a set). After I had all of my samples, I set them into the PCR machine, ran the “PAUL” PCR protocol and collected them when they were finished (Figure 2). Next week, I will be using the next set of primers: 27F and 1492R.

Figure 1. From left to right, PCAT-4r-2015, PCAT-4f-2015, 27F, 1492R and Tris Hydrochloride. The first 4 are the primers I will be working with and the last item is the solvent the primers needed to be dissolved in.

Figure 2. Collected samples after PCR was complete. The DNA samples used were from Group A (except for #12, which was from Group C).