Sample information

The collected specimen was a small insect, confirmed via DNA barcoding (COI gene sequencing) and BLAST analysis to be Cerodontha dorsalis (the Leaf-miner Fly). The identification showed a perfect 100.00% percent identity match.

This species belongs to the Order Diptera (True Flies) and is ecologically significant as an agricultural pest due to the larvae’s leaf-mining activity.

Testing for the endosymbiont bacterium Wolbachia using PCR and Gel Electrophoresis revealed no 438 bp band , concluding that this individual is Wolbachia-negative.

Methods

The DNA was successfully extracted from the Whole arthropod specimen (identified as Cerodontha dorsalis ) using a Qiagen DNeasy Blood and Tissue Kit with modifications.

PCR was performed as a Duplex Reaction to simultaneously amplify the COI gene (~700 bp) for species identification, and the Wolbachia gene (~438 bp)  for infection status.

The products were visualized via Gel Electrophoresis using a 1% Agarose Gel and 1X TAE buffer. Results confirmed the successful amplification of the COI gene (clear ~700 bp band) but showed no 438 bp band , indicating the specimen is Wolbachia-negative

Results

High Confidence. We can trust these results because several controls functioned properly.

1. Successful DNA Extraction & PCR: The COI gene was successfully amplified (clear band at ~700 bp) for the specimen. The COI gene serves as an internal positive control, confirming that the DNA extraction worked and the PCR mixture was functional.

2. High-Quality Sequencing: The sequencing process yielded high-quality base calls (Phred scores) , resulting in a 100.00% percent identity match via BLAST for the species ID.

3. Clear Negative Result: The expected band for Wolbachia infection was clearly absent 6, allowing for a definitive conclusion: the specimen is Wolbachia-negative