Sample information |
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| Picture |
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|---|---|
| Location | |
| Collection date | 09/08/2024 |
| Captive / Cultivated? | Wild-caught |
| Group | Bluegrass Community and Technical College |
| Observations | The specimen was collected using a flying insect trap containing banana peels and vinegar per the Arthropod Collection Traps Guide provided by the Wolbachia Project. The trap was hung from a tree branch adjacent to a small creek in a suburban backyard. The specimen was identified as an Aedes using the iNaturalist app. |
| Putative identification | Arthropoda Hexapoda Insecta Diptera Culicidae Aedes |
Methods |
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| Extraction kit | DNeasy (Qiagen) blood and tissue kit |
| DNA extraction location | Whole arthropod |
| Single or Duplex PCR | Duplex Reaction |
| Gel electrophoresis system | Standard electrophoresis system |
| Buffer | TBE |
| DNA stain | SYBR Safe |
| Gel images |
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| Protocol notes | GoTaqGreen polymerase was used. Reaction conditions were per Lab 3: PCR (Wolbachia Project Guide) for duplex reaction PCRs. The PCR reaction volume was 25 uL. 10 uL of each reaction was run on a 2% agarose gel. The relevant gel lane (lane 2) for the specimen #27 is boxed. |
Results |
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| Wolbachia presence | Yes |
| Confidence level | High |
| Explanation of confidence level | PCR products of ~440 bp (which is the correct size for the Wolbachia product) and ~700 bp (correct size for the CO1 product) were produced. All control reactions (Lanes 6 through 10) 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. |
| 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
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| Summary | The Aedes was found to be postive for Wolbachia. |
Water Boatman