Peach Root Weevil

November 20, 2023

Sample information
Picture
Photos by: Danielle P.
Location
Collection date 09/19/2023
Captive / Cultivated? Wild-caught
Group St. Albans School
Observations

This arthropod was found in a science classroom of the Saint Albans School in Washington, DC in early September.
Putative identification Arthropoda Hexapoda Insecta Coleoptera Curculionidae Pseudoedophrys Pseudoedophrys hilleri

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Whole arthropod
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniPCR
Buffer TBE
DNA stain GelGreen
Gel images
Protocol notes

Analysis: We had to rerun the arthropod DNA amplification for a second time to get the DNA to appear on a gel. The uploaded photo is from the second try, which turned out to be successful. In our first attempt, the DNA of the arthropod controls and of one other specimen were successfully amplified, but the DNA control and my specimen were not. We then reran PCR, but using a slightly larger quantity of DNA than we had the first time. This rerun produced the expected results.

 

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

Our controls all worked as expected, so we have no reason to doubt the outcome of gel electrophoresis. Additionally, another bug on the gel did test positive for Wolbachia, reenforcing that the gel was run properly.
Wolbachia 16S sequence
Arthropod COI sequence Download FASTA    Download AB1
GAAATTGACTANTACCTTTAATATTAGGAGCCCCAGATATAGCCTTTCCTCGACTAAATAATATAANATTTTGGCTTCTCCCTCCTTCTCTCTCCCTTCTATTANTAANAANAATTGTGGATAAAGGAGCAGGGACAGGATGAACCGTATACCCGCCCNTATCTGCTAATATCGCTCACGAAGGCTCATCTGTANACCTAGNTNTTTTCNNACTACATATAGCAGGAATTTCATCANTCCTANGAGNNNNNNNTTTTATTTCTACGGTAATTAATATACGTCCTTCANGAATATCATTTGATCGTTTACCTCTTTTTGTATGANCCGTTAAAATTACTGCTA
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Pseudoedophrys hilleri was found to be negative for Wolbachia.
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