Emily Wolbachia

September 27, 2021

Sample information
Picture
Photos by: Emily H.
Location
Collection date 09/02/2021
Captive / Cultivated? Wild-caught
Group Hampton Roads Academy
Observations

Red eyes, black or grey body, wings, furry on the body and legs.
Putative identification Arthropoda Hexapoda Insecta Diptera

Methods
Extraction kit Edwards Buffer
DNA extraction location Abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniOne
Buffer TBE
DNA stain GelGreen
Gel images
Protocol notes

When making the PCR master mix, we thought we made enough for 10 samples, even though we had 9. However, we needed to make more because we ran out for some reason. Later on, we accidentally unplugged the PCR machine when we were trying to find an outlet for the gel electrophoresis machine. We started the PCR again, but it did not turn off when it was supposed to. When we tried to turn on the gel electrophoresis machine, it did not work at all. We had to wait a couple days to get another one and then it finally worked.

Results
Wolbachia presence No
Confidence level Medium
Explanation of confidence level

There is no marker indicating that Wolbachia is present in the insect (03), but the line is a little blurred. The controls worked well, and the primer dimers showed up like they usually do. The controls worked on both the Wolbachia gel and the Arthropod gel. There was a band for the insect (03) in the arthropod gel.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Diptera was found to be negative for Wolbachia.
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