Mosquito 27

December 15, 2024

Sample information
Picture
Photos by: Mark E.
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 Insecta Diptera Culicidae Aedes

Methods
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
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
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
Arthropod COI sequence
The PCR product was not sequenced.
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Aedes was found to be postive for Wolbachia.
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