|Captive / Cultivated?||Wild-caught|
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|
|Extraction kit||DNeasy (Qiagen)|
|DNA extraction location||Partial abdomen|
|Single or Duplex PCR||Single Reaction|
|Gel electrophoresis system||MiniOne|
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.
|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.|