Sample information |
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| Picture |
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|---|---|
| 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 |
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| Extraction kit | DNeasy (Qiagen) |
| DNA extraction location | Abdomen |
| Single or Duplex PCR | |
| Gel electrophoresis system | MiniPCR |
| Buffer | TBE |
| DNA stain | GelGreen |
| Gel images |
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| 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 |
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| 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 |
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| Summary | The Coleoptera was found to be negative for Wolbachia. |
Water Boatman