Sample information

I found this Spider in my house near Washington, D.C. (which is kept at a temperature of approximately 72 degrees F). I collected it in the morning, around 7:30 AM, and the outdoor temperature that day was in the 70s (F). The spider is small/medium sized with red/brown coloring, long legs, and bulging circular abdomen. I later found it to be a pregnant female. It had low energy and did not move a lot. It also had a tendency to hide in dark areas and play dead when frightened.

Methods

DNA extraction: I made sure to crush the arthropod really well. I dissected the abdomen out, and did not include the exoskeleton.

Gel electrophoresis lanes for CO1 arthropod PCR:

1- 10kb DNA ladder.

2-The second well is my earwig (E-1)

3-The third well is my bee (E-2)

4- The fourth well is my spider (E-3)

5- The fifth well is my cricket (E-4)

6- The sixth well is my (+) arthropod control

7- The seventh well is my (-) arthropod control

8- The eight well is my DNA control

9- The ninth well is my H20 control

Analysis: My controls worked, but sample E-1 did not have a band. There could have been a problem with the primer sticking.

Gel electrophoresis lanes for Wolbachia PCR:

1- 10kb DNA ladder.

2-The second well is my earwig (E-1)

3-The third well is my bee (E-2)

4- The fourth well is my spider (E-3)

5- The fifth well is my cricket (E-4)

6- The sixth well is my (+) arthropod control

7- The seventh well is my (-) arthropod control

8- The eight well is my DNA control

9- The ninth well is my H20 control

Analysis: My controls worked, so I am confident that there was no contamination.

Results

All of my controls worked as expected. I did not have any problems with the protocol, and I was not surprised by any bands on the gel. Therefore, I am confident that my arthropod is not infected with Wolbachia.