Sample information

I found it flying around in my house.

Methods

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

The specimen was incubated for 30 minutes but checked every 15 minutes to vortex.

Our first PCR reaction was set up on 10/14/25 and was a duplex reaction because we used both the arthropod C01 and Wolbachia 16s primers together. Then used an annealing temperature of 49 degrees Celsius. we used this Taq polymerase: New England Biolab’s OneTaq Hot Start Quick load 2X Master Miz with Standard Buffer (#M0488S)

Our first gel image was taken on 10/21/25 and was run at 125 volts for 20 to 25 minutes, Used DNA Ladder: New England Biolabs 1 kb Plus DNA Ladder for Safe Stains (products # N0559S)

Our 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 F,R primers. Used an annealing temperature of 55 degrees Celsius.

Our second gel was taken on 11/4/25 and was run at 125 volts for 20 to 25  minutes. Used this DNA Ladder: New England Biolabs 1 kb Plus DNA Ladder for Safe Stains (products # N0559S)

Results

Gel #1 Both of my positive controls showed 2 bands.

Gel #2 All of our controls showed up at 438 bp which suggests cross contaminated. The water didn’t have a band, But the -C had a band which should not happen