Sample information

I found it on a window in warm environment indoors.

Methods

1. We used a DNA extraction protocol based on the insect adaption of New England Biolabs Monarch Spin gDNA Extraction kit(product# T3010)
2. The species was incubated for 30 minutes in a hot water bath at 56 degrees Celsius.

• 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 October 21, 2025 and:
• was run at 250 volts for 21 minutes
• used the DNA ladder: New England Biolabs 1 kb Plus DNA Ladder for Safe Stains(product # N0559S)
• • Our second PCR reaction was set up on 10/28/2025 , used the same Taq polymerase as the first PCR reaction, and:
  • was a single reaction that  wolbachia_F  primer(s).
  • used an annealing temperature of  55 degrees Celsius
• Our second Gel image was taken on 11/04/2025 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

Yes, the results for the four control lanes match expectations because the positive control also shows the correct band, while the negative control shows no bands. This means there was no contamination, so the experiment is valid.

We did our second round of gel electrophoresis for the single PCR set up on 10/29/2025 with the Wolbachia primers. Confidence level became even higher than before because we only did this with duplex, which was positive, and now single, which is also positive. The results for the four control lanes match expectations because the positive control also shows the correct bands, while the negative control shows no bands. This means there was no contamination, so the experiment is valid.