Sample information

Picture
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

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
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

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

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