Sample information

Found underneath a rock outside. Looks to have 50+ legs. I believe it is a millipede or a centipede. Appears to have 2 antennas. Brown, black, dark tan, and shiny. Eyes not visible. Found in terrestrial habitat and caught when the temperature was 74* F.

Methods

DNA Extraction:

Arthropod was easy to extract DNA from. Crushed easily. No mistakes recorded.

PCR:

We ran the PCR once and then we ran it again because we couldn’t see any bands for our arthropod gel electrophoresis. The second time the PCR went as planned. We did not identify any signs of contamination.

Gel electrophoresis, Arthropod CO1:

In the arthropod gel we did not find a band under the B-5 lane. In the B-5 and B-6 lanes of the gel electrophoresis, we had the corresponding DNA for each arthropod. In the W+ lane was a Wolbachia positive control and in the W- lane was a Wolbachia negative control, and in the H20 lane was a water control. We found bands for arthropod B-6 and our W+ and W- controls. Our controls worked meaning that something went wrong for this arthropod specifically. We cannot be certain about any results from this arthropod because there was not enough DNA to confirm that it was an arthropod.

Gel electrophoresis, Wolbachia:

There was not a band for Wolbachia in the B- 5 lane of our gel. In the B-5 and B-6 lanes of the gel electrophoresis, we had the corresponding DNA for each arthropod. In the W+ lane was a Wolbachia positive control and in the W- lane was a Wolbachia negative control, and in the H20 lane was a water control. We found a band for our W+ control. There was no band for arthropods B-6 and our W- control. Our controls worked as intended but there was not sufficient DNA for the arthropod gel so we cannot conclude if there was Wolbachia in our B-5 arthropod or not. We did not send the DNA to the lab for testing because there was no band present for the arthropod gel meaning it did not have enough DNA to get results from the lab.

Results

In the arthropod gel electrophoresis a band did not show up under the B-5 lane. This could be due to the primers not binding correctly or that enough DNA was not extracted from the arthropod. Because of this we cannot be sure if the arthropod was positive or negative for Wolbachia.