Mosquito 26

Sample information

Picture
Photos by: Mark E.
Location
Collection date 09/10/2024
Captive / Cultivated? Wild-caught
Group Bluegrass Community and Technical College
Observations

The specimen was discovered feeding on my leg. It was gently swatted and preserved in 70% alcohol until analysis.

The specimen was identified as an Aedes mosquito using the iNaturalist app.

Putative identification Arthropoda Hexapoda Insecta Diptera Culicidae Aedes

Methods

Extraction kit DNeasy (Qiagen) blood and tissue kit
DNA extraction location Whole arthropod
Single or Duplex PCR Single Reaction
Gel electrophoresis system Standard electrophoresis system
Buffer TAE
DNA stain Ethidium Bromide
Gel images
Protocol notes

GoTaqGreen polymerase was used for the PCR. 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, lanes 4 and 5, for the specimen (#26) are boxed.

Results

Wolbachia presence No
Confidence level High
Explanation of confidence level

No PCR product was obtained in the reaction using the Wolbachia primers (WO, Lane 4), while the reaction using the cytochrome oxidase 1 (CO, Lane 5) 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 (Lanes 10 through 15) 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
Arthropod COI sequence
The PCR product was not sequenced.
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Aedes was found to be negative for Wolbachia.
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