Alatopedes campestris A5

Sample information

Picture
Location
Collection date 09/28/2024
Captive / Cultivated? Wild-caught
Group Edmund Burke School
Observations

caught in a public park, temperature of about 70 degrees Fahrenheit, terrestrial habitat.

 

Two big grey wings, two antennae, appears to be a moth.

Putative identification Arthropoda Hexapoda Insecta Lepidoptera

Methods

Extraction kit DNeasy (Qiagen)
DNA extraction location Whole arthropod
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniOne
Buffer TBE
DNA stain GelGreen
Gel images
Protocol notes

DNA Extraction:

I made sure to crush the arthropod very well, but there were large chunks of exoskeleton that were difficult to break up. As a result, the sample had to be spun 1-2 extra times in the centrifuge for the steps using the filter.

 

Gel Electrophoresis, arthropod CO1:

1 – DNA ladder

2 – A4 DNA

3 – A5 DNA

4- A6 DNA

5 – Arthropod positive control

6 – Arthropod negative control

7 –  water

Analysis: all controls showed expected bands, and the sample showed an arthropod CO1 band.

Gel electrophoresis, Wolbachia:

1 – DNA ladder

2 – A4 DNA

3 – A5 DNA (lascoria ambigualis)

4- A6 DNA

5 – Wolbachia positive control

6 – wolbachia negative control

7 –  water

Analysis: all controls showed expected bands, and the sample did not show a wolbachia band.

 

Results

Wolbachia presence No
Confidence level High
Explanation of confidence level

All of my controls worked as expected. I did not have any problems
with the protocol, and I was not surprised by any bands on the gel.
Therefore, I am confident my arthropod is not infected with
Wolbachia.

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