Sample information

my arthropod was a Dark-winged Fungus Gnats its order is diptara its family is Cecidomyiidae the scientific name is Genus Afrotricha, it does have Wolbachia 

Methods

Gel electrophoresis system: Edvotek Gel Electrophoresis
Buffer: 1X TAE
DNA Stain: SYBR Safe

My first gel Image was taken on 10-21-25 and was run at 125 Volts for 30 minutes Using New England Biolabs 1 kb Plus DNA Ladder for Safe Stains (product # N0559S). This round told me that my arthropod tested false for Wolbachia

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.
We used both the Arthropod CO1 and Wolbachia 16S primer(s).

while using an annealing temperature of 55 degrees Celsius.

Second Round:

Gel electrophoresis system: Edvotek Gel Electrophoresis

Buffer: 1X TAE

DNA Stain: SYBR Safe

Protocol notes: ADD the following statements:

Our second Gel image was taken on 11-4-25 and:
was run at 125 volts for 30 minutes
used this DNA Ladder: New England Biolabs 1 kb Plus DNA Ladder for Safe Stains (product # N0559S)
Results: I am Highly positive that my arthropod is  Wolbachia infested, the reason for this is because after running the single Twice to check for any errors It came out with the same results witch leads me to Say in confidence that my arthropod test positively

This week we did DNA sequencing. For our arthropods only had the ZK-1 Come out due to the CD-1 arthropod being too distorted and messy.CO1: SequenceS16: Sequence 

Results

my confidence would have been high because i did 2 PCRs and both times mine came back positive,

however the BLAST results for the arthropod CO1 gene were a 100% match for Drosophila melanogaster, so it is likely that we mislabeled the sample and mixed it up with the positive control arthropod fruit fly.