GRP20 Housefly

May 24, 2024

Sample information
Picture
Photos by: Natalia F., Kira O., Ramon A., Elisa T., Mary M.
Location
Collection date 05/10/2024
Captive / Cultivated? Wild-caught
Group California Academy of Mathematics and Science
Observations

The housefly was caught outside on the rims of a trashcan.
Putative identification Arthropoda Hexapoda Insecta Diptera

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Whole arthropod
Single or Duplex PCR Duplex Reaction
Gel electrophoresis system Standard electrophoresis system
Buffer TAE
DNA stain Fast Blast
Gel images
Protocol notes

Wolbachia Gel Lanes (Left)
Wells 1-2: Empty
Well 3: DNA Ladder
Well 4: Arthropod (Winged) #1 – Fruit Fly
Well 5: Arthropod (Winged) #2 – Housefly
Well 6: Arthropod (Winged) #3 – Pale Beauty Moth
Well 7: Positive Arthropod Control
Well 8: Negative Arthropod Control
Well 9: Positive DNA Control
Well 10: Water Control
Wells 11-15: Empty

Arthropod Gel Lanes (Right)
Wells 1-2: Empty
Well 3: DNA Ladder
Well 4: Arthropod (Winged) #1 – Fruit Fly
Well 5: Arthropod (Winged) #2 – Housefly
Well 6: Arthropod (Winged) #3 – Pale Beauty Moth
Well 7: Positive Arthropod Control
Well 8: Negative Arthropod Control
Well 9: Positive DNA Control
Well 10: Water Control
Wells 11-15: Empty

We diluted 100x Fast Blast to 1X and left the gels to stain overnight on a rocker.

Results
Wolbachia presence Unknown
Confidence level High
Explanation of confidence level

No bands appeared on either gel so nothing can be said on whether or not the housefly had Wolbachia or not. There was mostly likely an error in both the DNA Extraction and the running of the gel.
Wolbachia 16S sequence
Arthropod COI sequence
Summary
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