Hello! This final week of the semester has been consumed by data collection and its requisite formatting into both the required research paper and oral presentation, but I did manage to find time in the lab to complete one last experiment. As mentioned in previous Semester 2 blogs, the primary focus this term has been on perfecting extraction protocols and electrophoresis gel production techniques. This focus was rewarded by repeated successful DNA extractions from the initial bacteria,
E. coli, and the project was also extended to included two additional gram-negative species,
P. aeruginosa and
P. vulgaris, as well as one gram-positive organism,
S. aureus.
As a final test before the next phase of my project, I took my successfully extracted samples and ran a PCR on them using my existing primers. I then tested the resultant samples for DNA using the gel technique I developed this semester. The result? Negative for DNA. However, this is not discouraging news. In fact, the results simply confirmed my suspicion that the primers I selected based on my original review of results published in
Critical Evaluation of Two Primers Commonly Used
for Amplification of Bacterial 16s rRNA Genes (Frank, et. al., 2008) would not work for the species in this study. This leads me directly to the next step of this project, designing my own primers, which will resume in January, 2015. Until then, cheers!
A scanning electron micrograph image of Pseudomonas aeruginosa cultured onboard shuttle mission STS-115, courtesy www.nasa.gov .
