Electrophoresis & PCR

Honestly, I am trying to stay hopeful because I already want to identify what species of bacteria I am dealing with, even though I am having a bit of a tough time doing so. However, science isn’t always perfect and adjustments have to be made to further in research, even if they’re small. With that said, patience is definitely key in this project. Similar to the past few weeks, I performed PCR and electrophoresis. I ran my post-PCR samples from last week in a gel and the results are shown below in Figure 1. These samples contained 1 microliter of DNA and 5 microliters of water and even though most of the slots showed DNA, there were a few that didn’t. I then decided to switch primers, from the PCAT set to the 27F/1492R set. There’s a possibility that the reason the DNA isn’t showing is because it doesn’t work in the PCAT primer. In figure 2 are post-PCR samples in the 27F/1492R primer set ran under electrophoresis. Luckily, slot 11 had DNA amplification but the others still weren’t successful. Next week, I will continue using the 27F/1492R primer set for PCR and double the amount of master mix, primers and sterile water to have a total volume of 40 microliters, instead of the usual 20. The amount of DNA in each PCR sample will stay at 1 microliter.

Figure 1. Electrophoresis results when adding post-PCR samples into each slot in the PCAT primer set. From left to right, the material in the slots are: MW Ruler, 1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14. The bands that are shown could possibly be the primers. 

Figure 2. Electrophoresis results when adding post-PCR samples into each slot in the 27F/1492R primer set. From left to right, the material in the slots are: MW Ruler, 1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and E. Coli. The only slot that showed DNA amplification was slot 11, which contained DNA from the unknown bacteria #12.