Sample information |
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| Picture |
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|---|---|
| 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 | Arthropoda Hexapoda Insecta Coleoptera Scarabaeidae Phyllophaga |
Methods |
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| 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 |
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| 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 and labeled the gels, it looks like there was some DNA that escaped from the gel- I did not use a loading dye, this is probably why. To stop this, I should add loading dye. |
Results |
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| Wolbachia presence | No |
| Confidence level | High |
| Explanation of confidence level | |
| Wolbachia 16S sequence | |
| Arthropod COI sequence |
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| Summary | The Phyllophaga was found to be negative for Wolbachia. |
Water Boatman