Willow Leaf Beetle

October 16, 2022

Sample information
Picture
Photos by: Daniel G., Adrian S.
Location
Collection date 09/20/2022
Captive / Cultivated? Wild-caught
Group Palm Beach Atlantic University
Observations

Black beetle with striped or dotted pattern on exoskeleton. The superior portion of the insect has a midline and was collected and stored in ethanol one day prior to experimentation.
Putative identification Arthropoda Insecta Coleoptera Chrysomelidae Chrysomela Chrysomela vigintipunctata

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

Two wild-caught arthropods were collected. A 6X loading dye was used for the gel and run at 100V with the power supply. The order of samples from lanes 1-7 are as follows: 100bp ladder, Arthropod #1 (Willow Leaf Beetle), Arthropod #2 (White weevil), positive control, negative control, positive gDNA, and a water control.

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

The protocol was followed and executed step by step with little to no error. For Lane 2, this simply gives us the insect DNA. The controls give a visual as to where the infected DNA would have ended if the insects did contain Wolbachia.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Chrysomela vigintipunctata was found to be negative for Wolbachia.
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