Tiger Crane Fly

Sample information

Picture
Photos by: Kaitlyn D.
Location
Collection date 09/18/2022
Captive / Cultivated? Wild-caught
Group Edmund Burke School
Observations

I found this tiger crane fly resting on a surface outside. I collected it at night, near midnight, and it was 67°F.

Putative identification Arthropoda Hexapoda Insecta Diptera Tipulidae

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 really well. I used the whole arthropod.

Gel Electrophoresis Lanes: For CO1 Arthropod PCR

1- 10Kb ladder

2- The second well is my first sample

3- The third well is my second sample

4- The fourth well is my third sample

5- The fifth well is my fourth sample

6- The sixth well is my negative  Arthropod DNA

7- The seventh well is my positive  Arthropod DNA

8- The eighth well is my positive Wolbachia DNA

9- The ninth well is water, and thus negative overall

Gel Electrophoresis Lanes: For Wolbachia Specific Gene  PCR

1- 10Kb ladder

2- The second well is my first sample

3- The third well is my second sample

4- The fourth well is my fourth sample

5- The fifth well is my third sample

6- The sixth well is my positive Arthropod DNA

7- The seventh well is my negative Arthropod DNA

8- The eighth well is my positive Wolbachia DNA

9- The ninth well is water, and thus negative overall

Results

Wolbachia presence No
Confidence level High
Explanation of confidence level

I’m confident in our results because the controls that were supposed to be positive were positive but our samples were not, indicating that our samples were not positive for Wolbachia.

Wolbachia 16S sequence
Arthropod COI sequence Download FASTA    Download AB1
Summary The Tipulidae was found to be negative for Wolbachia.
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