Sample information

The arthropod was caught as it was perched on a flower. It was taken on a field close to a stream. The animal was not harmed during the capture.

Methods

PCR reactions were set up on ice using Taq Master Mix, primers, water, template DNA, or controls to amplify the arthropod CO1 and Wolbachia genes. After that, the reactions were carried out in a thermal cycler. A 2% agarose gel was produced by melting 0.8g of agarose in 40mL of TBE buffer, adding 4µL of SYBR Safe dye, and allowing it to solidify on a casting tray. The gel was filled with TBE buffer and placed in an electrophoresis chamber with the wells facing the negative electrode. The amplified samples were then mixed with loading dye, loaded into the wells next to a DNA ladder, and run for 15 to 25 minutes so that the DNA bands could be seen under blue light.

Results

The strong, distinct band observed in the arthropod COI gene gel test demonstrated a very good DNA extraction using the DNeasy kit. The other gel test for the 16S Wolbachia gene also revealed a distinct, bright band in the appropriate lane for this specimen, providing strong visual evidence of a positive result.