Sample information

The specimen was collected using a flying insect trap containing banana peels and vinegar per the Arthropod Collection Traps Guide provided by the Wolbachia Project. The trap was hung from a tree branch adjacent to a small creek in a suburban backyard for two days. The trap was cut open and the (predominantly flying) insects were removed and preserved in 70% alcohol. The trap was then placed on the ground for an additional three days capturing additional insects. These insects included both crawling and flying insects. These insects were removed and preserved in 70% alcohol. 

The specimen was identified as a Diptera using the iNaturalist app

Methods

GoTaqGreen polymerase was used. Reaction conditions were per Lab 3: PCR (Wolbachia Project Guide) for single reaction PCRs (separate reactions were run with the Wolbachia primers and the cytochrome oxidase primers). The PCR reaction volume was 25 uL. 5 uL of each reaction was run on a 2% agarose gel. The relevant lanes, lanes 1 and 2, for the specimen (#8) are boxed.   

Results

No PCR product was obtained in the reaction using the Wolbachia primers (WO, Lane 1), while the reaction using the cytochrome oxidase 1 (CO, Lane 2) produced a ~700 bp product, which is the correct size for the CO1 product, indicating the DNA isolated from the specimen is of suitable quality for PCR. All control reactions (Lanes 9 through 14) worked as they should. The positive control is DNA extracted from a control insect infected with Wolbachia, while the negative control is DNA extracted from an uninfected insect. Both controls were provided by the Wolbachia Project.