Sample information |
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| Picture |
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|---|---|
| Location | |
| Collection date | |
| Captive / Cultivated? | Wild-caught |
| Group | Bluegrass Community and Technical College |
| Observations | Caught in backyard of residence. The specimen was identified as a stink bug using the iNaturalist app. |
| Putative identification | Arthropoda Hexapoda Insecta Hemiptera Pentatomidae |
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 | Standard electrophoresis system Bio-Rad |
| Buffer | TAE |
| DNA stain | Ethidium Bromide |
| Gel images |
|
| Protocol notes | GoTaqGreen polymerase was used. Reaction conditions were per Lab 3: PCR (Wolbachia Project Guide) for single reaction PCRs (separate reactions were run with the Wolbachia primers and the cytochrome oxidase primers). The PCR reaction volume was 25 uL. 5 uL of each reaction was run on a 2% agarose gel. The relevant lanes for the specimen are boxed in the gel picture. |
Results |
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| Wolbachia presence | No |
| Confidence level | High |
| Explanation of confidence level | No PCR product was obtained in the reaction using the Wolbachia primers while the reaction using the cytochrome oxidase 1 produced a ~700 bp product, which is the correct size for the CO1 product, indicating the DNA isolated from the specimen is of suitable quality for PCR. All control reactions worked as they should. The positive control is DNA extracted from a control insect infected with Wolbachia, while the negative control is DNA extracted from an uninfected insect. Both controls were provided by the Wolbachia Project.
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| Wolbachia 16S sequence |
The PCR product was not sequenced.
BLAST at The Wolbachia Project BLAST at NCBI
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| Arthropod COI sequence |
The PCR product was not sequenced.
BLAST at The Wolbachia Project BLAST at NCBI
|
| Summary | The Pentatomidae was found to be negative for Wolbachia. |
Water Boatman