Spotted Cucumber Beetle

October 9, 2020

Sample information
Picture
Photos by: Elijah D.
Location
Collection date 08/14/2020
Captive / Cultivated? Wild-caught
Group St. John High School USD 350
Observations

I found these Spotted Cucumber beetles on sunflowers. Before I preserved them in 70% ethanol they were a bright yellow and they blended in with the flowers really well. There were also Soldier beetles on the same sunflowers and blended in as well.
Putative identification Arthropoda Hexapoda Insecta Coleoptera

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

The beetle was stored in 90% ethanol for about a month before extraction.

DNA Extraction: The DNA was extracted from the abdomen. All of the Dneasy (Qiagen) protocols were followed. The extraction took several days to complete.

PCR: Mini One Taq polymerase was used.

Gel Electropheresis: All of the lanes worked except for the DNA ladder. When pipetting, the well was missed and had to be done again which might have been the reason.

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

All of the controls worked and were very clear. The DNA extraction protocols were followed exactly.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Coleoptera was found to be negative for Wolbachia.
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