May beetle

Sample information

Picture
Photos by: Athena L.
Location
Collection date 04/08/2021
Captive / Cultivated? Wild-caught
Group Bordenstein Lab
Observations

We noticed these beetles that would come out every night around 8pm and bother us. They were loud and would run into us. Not very intelligent, they would routinely get stuck on their backs and would not be able to do anything until they fell off the surface they were on. There were so many of them! We thought they had something to do with the grape plant next to our fence, grape beetles look very similar but have spots. These have no spots. We identified them as lawn grubs, and these are May/June beetles.

Putative identification Hexapoda Insecta Coleoptera Scarabaeidae Phyllophaga

Methods

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

DNA extraction: Followed protocol, removed exoskeleton and extracted goop from inside the bug. I tried to get as much of the white goop as possible, and as little of the exoskeleton.

No problems with the PCR protocol. I used the primers from The Wolbachia Project. We ran the arthropod PCR with an annealing temperature of 49C, and the Wolbachia PCR with an annealing temperature of 55C.

 

I added both the arthropod and Wolbachia labelled gel. Everything looks good, but there is some smearing on the gel. I suspect this is because I did not use any loading dye, so some of the DNA came out of the well and floated around. I can prevent this next time by using loading dye for my samples.

Results

Wolbachia presence No
Confidence level High
Explanation of confidence level

I am confident in my results because my controls looked as expected. I had two bands for both of my positive controls, only a band in the arthropod control for my negative fruit fly control, and absolutely nothing in my negative water control.

I am confident there is not Wolbachia in this sample because there was no band in the lane that my sample was in visualizing the PCR product of my Wolbachia PCR.

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