Sample information

• The specimen been caught under a 5 foot long rock, under moist dirt in a garden flower.  Only odd thing that there was two stone centipedes that were around 8 inches near each other.

Methods

We used the DNA extraction protocol based on insect adaptation of new england’s biolabs monarch spin gDNA Extraction kit. (Product # T3010) Specimen was incubated for 15 minutes in a hot water bath at 56 degrees C.

First pcr was done on oct 14th 2025 and was a duplex reaction since both arthropod CO1 and wolbachia 165 prime together. Annealing temperature at 49 degrees Celsius. Used Taq polymerase new england biolabs hot start quick-load 2x master with standard buffer (M0488S) Our first gel was taken on 10/21/2025 and ran at 100   Volts for 25 minutes and Used DNA ladder:new england biolabs 1kb plus DNA ladder for safe stains (product # No559s)

Our third  PCR was a single  PCR that only had Woolbachia primers using the same equipment. Our third  gel image was taken on 11/4 was run at 125 volts for 25 minutes used DNA ladder: new england biolab 1kb plus DNA ladder safe for stains (product # No559s) 

Results

My current confidence level is at a medium for the two uncontaminated Duplex PCR showed zero Infected wolbachia inside the centipede. But It isnt a full 100 percent for because my single PCR for wolbachia failed due to a contamination. For full  condifence  a successfully  doing a single Wolbachia PCR would help solidy that there isnt any Wolbachia DNA whatsoever.