Sample information

Arthropod was collected outdoors in a dirt area.

Methods

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 first PCR was set up on 10/14/25 and was a duplex reaction because we used both the arthropod CO1 and Wolbachia 16S Primers together. Used an annealing temperature of 49 degrees Celsius. Used this TAQ polymerase: New England Biolabs OneTaq Hot start Quick Load 2X Master Mix with standard buffer  (#M0488S).
Differed from the written protocol: used 88 ul of Taq master mix

Our first gel image was taken on 10/21/25 and was run at 125 volts for 22 minutes. Used this DNA Ladder : New England Biolabs 1 kb Plus DNA Ladder for safe stains (product #N0559S)

Our second PCR reaction was set on 10/28/25 used the same Taq polymerase as the first PCR reaction, and was a single reaction that used Wolbachia F and R primers. Used an annealing temperature of 55 degrees Celsius.
Differed from the standard protocol in that : used 88uL instead of 87.5

Our second gel image was taken on 11/4/25 and was run at 125 volts for 27 minutes.
used this DNA ladder: New England BioLabs 1kb plus DNA Ladder for Safe Stains (product # N0559S)

Results

Confidence level will increase with more testing to prove accurate results. Controls matched expectations, except +arthropod control was not detected at all. My arthropod had a band around 700bp but no band for Wolbachia.

11/4/25 Confidence level for the second gel electrophoresis is high moderate. My arthropod still detected no Wolbachia like the previous one, but the +arthropod control still did not show any bands present. Confidence will increase as more testing is done.