Sample information |
|
| Picture |
|
|---|---|
| Location | |
| Collection date | 09/12/2023 |
| Captive / Cultivated? | Wild-caught |
| Group | Palm Beach Atlantic University |
| Observations | I found this mosquito near an alleyway between Palm Beach Christian Academy and Rinker School of Business in the afternoon, about 2:00 pm, in 87 degrees F. I attached a picture of when the mosquito was contained in a petri dish. |
| Putative identification | Arthropoda Hexapoda Insecta Diptera Culicidae |
Methods |
|
| Extraction kit | DNeasy (Qiagen) |
| DNA extraction location | Whole arthropod |
| Single or Duplex PCR | Single Reaction |
| Gel electrophoresis system | MiniPCR |
| Buffer | TAE |
| DNA stain | GelGreen |
| Gel images |
|
| Protocol notes | DNA extraction: made sure samples are crushed well and entire body used Used 2% agarose gel Used 4 uL of 6X loading dye Gel electrophoresis lanes: 1 – 10 uL 1 Kb Plus ladder 2 – (no sample) 3 – 20 uL of arthropod #1 (mosquito) 4 – 20 uL of arthropod #2 (ant) 5 – 20 uL of positive arthropod control 6 – 20 uL of negative arthropod control 7 – 20 uL of positive DNA control 8 – 20 uL of water Analysis: controls worked and extraction done properly because bands for arthropod gene are seen and all three control bands were as expected. Lane 8 is negative control with just water therefore no bands. |
Results |
|
| Wolbachia presence | No |
| Confidence level | High |
| Explanation of confidence level | All controls were as shown as expected results. No conflicts encountered during lab procedure. The size of arthropod CO1 gene is calculated to be 764 bp and Wolbachia band of 409 bp was not detected in the mosquito. |
| Wolbachia 16S sequence | |
| Arthropod COI sequence |
|
| Summary | The Culicidae was found to be negative for Wolbachia. |
Water Boatman