Carpenter Bee

November 22, 2022

Sample information
Picture
Photos by: Kaitlyn D.
Location
Collection date 09/07/2022
Captive / Cultivated? Wild-caught
Group Edmund Burke School
Observations

I found this carpenter bee while resting on a flower outside. I collected it in the afternoon, and it was 79°F.
Putative identification Arthropoda Hexapoda Insecta Hymenoptera Apidae Xylocopa

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniOne
Buffer TBE
DNA stain GelGreen
Gel images
Protocol notes
DNA extraction: I made sure to crush the arthropod really well. I dissected the abdomen out.

Gel Electrophoresis Lanes: For CO1 Arthropod PCR

1- 10Kb ladder

2- The second well is my first sample

3- The third well is my second sample

4- The fourth well is my third sample

5- The fifth well is my fourth sample

6- The sixth well is my negative Arthropod DNA

7- The seventh well is my positive Arthropod DNA

8- The eighth well is my positive Wolbachia DNA

9- The ninth well is my overall negative

Gel Electrophoresis Lanes: For Wolbachia Specific Gene  PCR

1- 10Kb ladder

2- The second well is my first sample

3- The third well is my second sample

4- The fourth well is my fourth sample

5- The fifth well is my third sample

6- The sixth well is my positive Arthropod DNA

7- The seventh well is my negative Arthropod DNA

8- The eighth well is my positive Wolbachia DNA

9- The ninth well is water, and thus negative overall

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

I’m confident in our results because the controls that were supposed to be positive were positive but our samples were not, indicating that our samples were not positive for Wolbachia. Unfortunately, the earwig did not have a bright enough band from the gel electrophoresis to have the DNA sequenced.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Xylocopa was found to be negative for Wolbachia.
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