Coleoptera C-2

October 31, 2024

Sample information
Picture
Photos by: Sam H.
Location
Collection date 09/17/2024
Captive / Cultivated? Wild-caught
Group Edmund Burke School
Observations

Caught in science lab in late summer. Spotted, orangeish color, slightly damaged wing.
Putative identification Arthropoda Insecta Coleoptera Coccinellidae

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Whole arthropod
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniOne
Buffer TBE
DNA stain GelGreen
Gel images
Protocol notes

DNA Extraction: Arthropod was crushed easily and there were no issues throughout the extraction process.

PCR: No mistakes throughout process.

Gel electrophoresis, Arthropod CO1: Well C2 was the Coleoptera, W+ and W- were positive and negative for Wolbachia but still contained arthropod DNA, and H2O is water to be a control for contamination. C2 had a band between 800 and 600 base pairs, same with W+ and W-. Water had no band so there is no reason to assume the samples were crosscontaminated.

Gel electrophoresis, Wolbachia: There was no band present for the C-2 lane indicating it was not positive for Wolbachia. We have no reason the believe this is false because the positive Wolbachia control and a band and the negative Wolbachia control and the the water control did not have bands.

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

The negative Wolbachia control had no band so we have no reason to believe the sample was contaminated.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Coccinellidae was found to be negative for Wolbachia.
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