Sample information

The arthropod was found under a rock initially rolled into a ball. Once the specimen was submerged into alcohol it unrolled.

Methods

• We had used a DNA extraction protocol based on the insect adaptation of New England Biolabs’ Monarch Spin gDNA kit (Product # T3010).
• The specimen was incubated for 30 minutes in a hot water bath at 56 degrees C.
• During our lab experiement, we had made one accidental mistake that made no difference in our results, but we had dropped our spin columns in the discard thinking we didnt need them and later realized we needed them for the end part of DNA Elution.
• Our PCR reaction was set up on 10/16/25 and: was a duplex reaction because we used both the Arthropod CO1 and Wolbachia 16S primers together.
• Taq Polymerase: New England Biolabs OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer (#M0488S) was used.
• Annealing temperature of 49 degrees Celsius.
• Our first Gel image was taken on 10/23/25 and: was run at 125 volts for 30 minutes.
• New England Biolabs 1 kb Plus DNA Ladder for Safe Stains (product # N0559S).
• Our second PCR Reaction was set up on 10/30/25, used same Taq Polymerase as the first PCR reaction:
• was a duplex reaction that used the Wolbachia and Arthropod primers.
• used annealing temperature of 49 degrees Celsius.
• Our second Gel image was taken on 11/6/25 and was run at 125 volts for 30 minutes.
• Used this DNA Ladder: New Biolabs 1kb Plus DNA Ladder for Safe Stains (product #N0559S).

Results

In our first Gel image it shows in row 2 (3-AT) negative results. There would be two bands if positive, but there are no bands.

In our second gel image it shows row 2(3-AT) has positive results for Wolbachia because it had a band at about 400bp and no bands for Arthropod.