Wolbachia #1: Mosquito

March 30, 2023

Sample information
Picture
Photos by: Kamilia B.
Location
Collection date 09/14/2022
Captive / Cultivated? Wild-caught
Group Woburn Memorial High School
Observations

– mosquito had a long physique and long legs
– a large abdomen
– made identifying the sex organs more difficult
Putative identification Arthropoda Hexapoda Insecta Diptera Culicidae

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

– cut off abdomen of mosquito in order to use the sex organs to find Wolbachia

– Cell lysis the DNA of the insect and then precipitate the DNA

– Then drain the DNA pellets in absorb the supernatant for PCR testing

– after running PCR use the DNA to run a 2% agarose gel electrophoresis

– Well 1: Ladder (100 bp)
– Well 2: arthropod #1 mosquito
– Well 3: arthropod #2 pill bug
– Well 4: (+) Arthropod Control
– Well 5: (-) Arthropod Control
– Well 6: (+) DNA Control
– Well 7: (-) DNA Control (Water)
– Well 8: nothing

– compare results with control DNA in order to identify wolbachia in the insect

Results
Wolbachia presence Yes
Confidence level Medium
Explanation of confidence level

I believe that there is no Wolbachia is in the mosquito. It is clear that there is no Wolbachia band below the arthropod band. However there was an indication of cross contamination, meaning not all results may be accurate
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Culicidae was found to be postive for Wolbachia.
Report Inappropriate Post