Sample information

The fly was found inside a screened in porch. It was flying around the porch and kept landing on the screen.

Methods

1.We used a DNA extraction protocol based on the insect adaptation of New England Biolabs’ Monarch Spin gDNA Extraction Kit(Product # T3010).

2. The specimen was incubated for 30 minutes in a hot water bath at 56 degrees C

• Our first PCR reaction was set up on October 14, 2025 and:

•was a duplex reaction because we used both Arthropod CO1 and Wolbachia 16S primers together

•used an annealing temperature of 49 degrees Celsius.

•used the Taq polymerase: New England Biolabs OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer(#M0488S)

• Our first Gel image was taken on 10/21/25 and: was run at 250 volts for 21 minutes .
• Used this DNA Ladder: New England Biolabs 1 kb Plus DNA Ladder for Safe Stains(product # N0559S)

• ur second PCR reaction was set up on 10/28/25 , used the same Taq polymerase as the first PCR reaction, and: 
• was a single reaction that used, the wolbachia primer(s).
• used an annealing temperature of 55 degrees Celsius.

Our second gel image was taken on 11/4/25 and: was run at 125 volts for 25 minutes.

• used this DNA ladder: New England Biolabs 1 kb Plus DNA Ladder for Safe Stains (product # N0559S).

Results

I am very confident the experimental results are valid. This is because my bands for 1-MO and 1-SB are very clear and obvious. I also am confident because my results for my control variables presented like they were supposed to. This shows that any data received is accurate.

For our second gel electrophoresis my confidence level for my arthropod being Wolbachia positive is very confident. I got the same exact results as my first gel electrophoresis except for I only did a single PCR targeting the Wolbachia primers. My controls also showed up how they were supposed to, therefore my data received is accurate.