Antelope Beetle

November 7, 2023

Sample information
Picture
Photos by: Norvana H., Deonte D.
Location
Collection date 11/01/2023
Captive / Cultivated? Wild-caught
Group John Overton High School
Observations

Most of the insects collected, specifically beetles, were found under logs; so, dark, small, and hidden places.
Putative identification Arthropoda Insecta Coleoptera

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Abdomen
Single or Duplex PCR Duplex Reaction
Gel electrophoresis system Standard electrophoresis system
Buffer TAE
DNA stain SYBR Safe
Gel images
Protocol notes

DNA Extraction: I took great care to thoroughly crush the arthropod, as well as removing the abdomen, leaving the exoskeleton behind.

Gel Electrophoresis Lanes for CO1 arthropod PCR: 

  1. DNA Ladder
  2. Arthropod #1
  3. Arthropod #2
  4. Arthropod #3
  5. Arthropod #4
  6. 6th well is the (+) Arthropod Control
  7. 7th well is the (-) Arthropod Control
  8. 8th well is the (+) DNA Control
  9. Finally, the 9th well is the sterile water

Results
Wolbachia presence No
Confidence level Medium
Explanation of confidence level

After receiving the results from the samples collected and tested, the bands for my collected sample, Arthropod #3, weren’t visible for Wolbachia. On the other hand, the bands for Arthropod #4 and the (+) Arthropod Control were both visible for Wolbachia.

The controls did show signs of success, still, I’d rather consult with scientists who were present before I come to a solid conclusion.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Coleoptera was found to be negative for Wolbachia.
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