Week 8: PCR and Large-scale DNA Extraction

This week was filled with results! I have had great success with the Nitric Acid digestions, and the lab now has permanent slides for the chironomid gut contents from June and July for all three collection sites! This is great because these results will be compared to Claudia's counts of the same sites to see how the environment compares to the chironomid diet. 

I also performed my first PCR on my original DNA extraction. We used primers that search for diatom DNA. Using these in the PCR and gel electrophoresis procedures, I was able to determine that there were diatoms inside the chironomid gut! This was a very exciting result because I had previously been unsure of my ability to extract DNA from organisms/tissues inside the chironomid. Because we saw bands for diatoms, I now know that we can test for the presence or abscence of various things in the diatom guts using our current extraction procedure. Very exciting! 

Because I got such a great result, I moved forward with DNA extractions from all nine sites, as well as controls. Unfortunately, the DNA was contaminated, likely by one of the elution buffers, and so it was not really usable. 

I finished the week by cleaning chironomids for another extraction on Monday. Hopefully that one will go better, and we can continue to do some PCR analysis of the chironomid gut contents.