Sample information |
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Picture |
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Location | |
Collection date | 03/31/2025 |
Captive / Cultivated? | Wild-caught |
Group | FernLeaf Community Charter School |
Observations | Triangular abdomen with mottled color, brown with white flecks. |
Putative identification | Arthropoda Hexapoda Insecta Hemiptera |
Methods |
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Extraction kit | DNeasy (Qiagen) blood and tissue kit |
DNA extraction location | Abdomen |
Single or Duplex PCR | Single Reaction |
Gel electrophoresis system | MiniOne |
Buffer | TBE |
DNA stain | GelGreen |
Gel images |
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Protocol notes | We removed the abdomen of our Halymorpha halys, then placed it into a centrifuge tube. We ground it up before adding buffer ATL, Proteinase K, and Buffer AL to the mixture. It was incubated for a week, then spun through a centrifuge and debris was removed. We added ethanol. then transfered it to a spin column. We centrifuged it for 1 minute, then added buffers AW1 and AW2. We moved it back to a microcentrifuge tube, then incubated for 1 hour after adding Buffer AE to the sample. We added a primer, then ran it through a Mini one gel electrophoresis machine. |
Results |
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Wolbachia presence | No |
Confidence level | Medium |
Explanation of confidence level | Conducted as a school experiment. Potential cross contamination between groups and bugs, although unlikely because of a clean negative control sample. PCR gel was stabbed during loading, which could’ve obscured or produced incorrect results. |
Wolbachia 16S sequence | |
Arthropod COI sequence |
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Summary | The Hemiptera was found to be negative for Wolbachia. |