Sample information

Was collected at a window in my house using toilet paper to capture them without crushing them, then putting them in a Ziplock bag full isopropyl alcohol to preserve until DNA extraction proceeded.

The insect was identified as Genus Halyomorpha 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 2 and 3, for the specimen (#17) are boxed.  

Results

No PCR product was obtained in the reaction using the Wolbachia primers (WO, Lane 3), 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 10 through 15) 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.