Beetle

November 9, 2023

Sample information
Picture
Photos by: Zaniya CC
Location
Collection date 11/01/2023
Captive / Cultivated? Wild-caught
Group John Overton High School
Observations

This is the environment the beetle was caught in.
Putative identification Arthropoda Hexapoda 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: Seeing that our controls don’t showcase any signs of DNA, I believe that part of the reason our arthropods don’t have any Wolbachia in them is because we didn’t crush them enough to extract the DNA.

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

The bands for my collected sample, Arthropod #2 weren’t visible for Wolbachia which led me to believe that we did a poor job at crushing our arthropods up in the very beginning. This made our results inaccurate.

 
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Coleoptera was found to be negative for Wolbachia.
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