Sample information

It was outside near the corner of the building in moist dirt near leaves and rocks.

Methods

Our first PCR reaction was set up on 10/14/25 and was a duplex reaction because we used both the Arthropod CO1 and Wolbachia 16S primes together. We used an annealing temperature of 49 degrees celsius. We used this Taq polymerase: New England Biolabs OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer (#M0488S).

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

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

Our second PCR reaction was set up on 10/28, we used the same Taq polymerase as the first PCR reaction and it was a duplex reaction that used Arthropod C01 and Wolbachia 16S primers. We used an annealing temperature of 49 degrees celsius.

Our second gel image was taken on 11/4/25 and was run at 125 volts for 25 minutes. We used the New England Biolabs 1 kb Plus DNA Ladder for safe stains (product #N0559S)

Results

My arthropod tested positive for Wolbachia, but it did not have a band for arthropod DNA. Therefore, I am not confident in the results.

I ran another duplex PCR, and my results showed nothing for Wolbachia. There was also no band showing any arthropod DNA, so I am confident that something went wrong in the beginning, maybe the DNA extraction.