Fly Wolbachia DNA

September 7, 2023

Sample information
Picture
Location
Collection date 11/03/2020
Captive / Cultivated? Wild-caught
Group Hampton Roads Academy
Observations

Has two wings and is green underneath. A little bit stripped on top. 4 legs.
Putative identification Arthropoda Hexapoda Insecta Diptera

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

-for DNA extractions, after I put the partial abdomen into the micro centrifuge I accidentally added 150 microlitters instead of 100

-for PCR steps, when making the master mix we thought we made enough for 10 (we had 9), but we had to made a little bit more because it ran out for some reason.

-our PCR machine did not work and we had to push experiment back a few days. Everything was stored safely and at correct temp and we made new gel on day of experiment.

-we did not need to add much green dye because our DNA was already died a couple days before. However in order to made it easier to pick up with a micro centrifuge we added 5 micro liters of green dye.

Results
Wolbachia presence Yes
Confidence level Medium
Explanation of confidence level

The band is pretty faint, this could be because of too much edwards buffer.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Diptera was found to be postive for Wolbachia.
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