Fruit fly (Drosophila melanogaster)

Sample information

Picture
Photos by: Renee Z
Location
Collection date 10/02/2025
Captive / Cultivated? Wild-caught
Group Berkshire Community College
Observations

Residential backyard, near some blueberry bushes and a tree stump. By a garden with some rotting tomatoes on the vine.

Putative identification Arthropoda Insecta Diptera Drosophilidae Drosophila

Methods

Extraction kit Monarch DNA extraction (NEB)
DNA extraction location Whole arthropod
Single or Duplex PCR Duplex Reaction
Gel electrophoresis system Edvotek Gel Electrophoresis
Buffer 1X TAE
DNA stain SYBR Safe
Gel images
Protocol notes

We used a DNA extraction protocol based on the insect adaptation of New England’s biolabs’ Monarch Spin gDNA Extraction kit.

the specimen was incubated for 30 minutes in a hot water back at 56 degrees C.

Our first Gel image was taken on 10/23/2025. It was run at 125 volts for 29 minutes. The DNA Ladder used was New England Biolabs 1kb Plus DNA Ladder for Safe Stains (product#N0559S

We ran a second PCR  – this time a single PCR and not a duplex – on 11/5/2025. A second gel electrophoresis was run on 11/6/2025. It was run at 125 volts for 25 minutes. The DNA Ladder used was New England Biolabs 1kb Plus DNA Ladder for Safe Stains (product#N0559S).

Results

Wolbachia presence No
Confidence level High
Explanation of confidence level

My confidence level is high that my arthropod was tested negative for Wolbachia because all other control samples were clear and accurate. The color and brightness for my and my partners arthropod to have the Arthropod DNA line visible but not Wolbachia line was very clear and obvious.

The second gel was ran from a PCR with Arthropod primer and not Wolbachia primer. The intention was to try a different method since my confidence was very high no Wolbachia was present.

Wolbachia 16S sequence
Arthropod COI sequence
Summary The Drosophila was found to be negative for Wolbachia.
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