B-3 (Hymenoptera)

November 21, 2025

Sample information
Picture
Location
Collection date 09/09/2025
Captive / Cultivated? Wild-caught
Group Edmund Burke School
Observations

Location Description: Specimen collected as it was landing on a flower in an open grassy field.

Season: Autumn

Temperature: 75 Fahrenheit

Habitat: Terrestrial

Notes: Yellowish and black hue, two wings, six legs, and two short antennae.
Putative identification Arthropoda Insecta Hymenoptera

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: No errors made. Changed pipette tips between each sample.

PCR: Pipette tip still held a bit of DNA solution after depositing 2 uL into the B-3 0.2 mL PCR Tube.

Gel electrophoresis, Arthropod CO1: In Lane 1 is a DNA Ladder, in Lane 2 is Sample #1, in Lane 3 is Sample #2, in Lane 4 is Sample #3, in Lane 5 is Sample #4, in Lane 6 is a Positive Wolbachia Control, in Lane 7 is a Negative Wolbachia Control, and in Lane 8 is Water. No errors were made during this procedure and controls worked as intended.

Gel electrophoresis, Wolbachia: In Lane 1 is a DNA Ladder, in Lane 2 is Sample #1, in Lane 3 is Sample #2, in Lane 4 is Sample #3, in Lane 5 is Sample #4, in Lane 6 is a Positive Wolbachia Control, in Lane 7 is a Negative Wolbachia Control, and in Lane 8 is Water. No errors were made during this procedure and controls worked as intended.

Results
Wolbachia presence Yes
Confidence level High
Explanation of confidence level

No mistakes were made during our gel electrophoresis and the controls for our Wolbachia and Arthropod CO1 gels were consistent with expected results, giving our sample results confidence.
Wolbachia 16S sequence Download FASTA    Download AB1
Arthropod COI sequence Download FASTA    Download AB1
Summary The Hymenoptera was found to be postive for Wolbachia.
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