Sample information

Collected on the shore of the Owego Creek

Methods

The Wolbachia gel electrophoresis protocol depicted in Figure. 1 shows no bands in any other column besides the DNA ladder.  There could be many explanations to why this part of the project failed, but a possibility could be that there was not enough DNA control left so we may have not received an accurate measurement of control affecting our results.

Results

The DNA ladders on both gel images were visible meaning the PCR machine ran correctly. The gel in figure 2 gave us a high level of confidence because it revealed all expected bands for the Arthropod Tubes. Figure 1 did not reveal the positive Arthropod or DNA control groups in the gel. There were no bands in either water control, leaving high confidence of no contamination.

Summary:

Figure 1. showed no bands of Wolbachia DNA or positive controls. In wells 5 and 7, there were no bands of DNA, suggesting that there was an error in our procedure. When extracting the positive DNA controls from our class sample, there might have not been enough material left, causing our measurements to be off. When initially crushing our arthropod samples, we might not have completely released DNA, leaving an inadequate amount of sample for the rest of our procedure. Other sources of error may have resulted from issues when transferring material via micropipettes.

Figure 2. presented bands for the DNA ladder, arthropod #1, arthropod #2, positive arthropod control, negative arthropod control, and positive DNA control. Our defined DNA ladder shows that our gel electrophoresis setup was functioning properly. The arthropod #1 & #2 bands show a clear presence of DNA extraction. The positive and negative arthropod controls were less defined, however this is most likely because the DNA was less concentrated. Our positive DNA control also showed a bright band while our negative DNA control did not, suggesting that there was no significant source of contamination.