Armadillidium nasatum

October 30, 2024

Sample information
Picture
Photos by: Alex D
Location
Collection date 09/04/2024
Captive / Cultivated? Wild-caught
Group St. Albans School
Observations

The mosquito was found in a leafy area near an oak tree in Rock Creek Park.
Putative identification Arthropoda Hexapoda Insecta Diptera Culicidae

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

DNA extraction: I made sure to crush the arthropod very well. Then, I dissected the abdomen and did not include the exoskeleton.

All of the lanes are labeled on the gel image

Lanes 9 and 10 are reruns of an originally unsuccessful attempt for a classmate’s Pill Bug

The Wolbachia bands were 438bp each

The CO1 bands were 708bp each

On the CO1 gel, my arthropod is labeled, along with the positive CO1 control and the 100bp ladder.

Results
Wolbachia presence Yes
Confidence level Medium
Explanation of confidence level

It is clear that the arthropod this sequence corresponds to has Wolbachia. However, we did not get a match for the same arthropod that I extracted DNA from (mosquito). The arthropod that we got from the sequence of nucleotides was a Roly Poly bug. This could have been a contamination error during the extraction or transportation process.
Wolbachia 16S sequence Download FASTA   
GCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCATCCTTAGTnGCnATCAGGTAATGCTGAnTACTTTAAGGAAACTGCCAGTGATAAGCTGGAGGAAGGTGGGGATGATGTCAAGTCATCATGGCCTTTATGGAGTGGGCTACACACGTGCTACAATGGTGTCTACAATGGGTTGCAAnGTGCGCAAGCCTAnnCTAnTCCCTAAAAGACATCTCAGTTCGGATTGTACTCTGCAACTCGAGTACnTGAAGTTGGAATCGCTAGTAATCGTGGATCAGCATGCCACGGTGAATACGTTCTCGGGTCTTGTACACACTGCCCGTCACGCCATGGGAATTG
BLAST at The Wolbachia Project   BLAST at NCBI
Arthropod COI sequence Download FASTA   
CTGGGAGTCTCATTGGGGATGATCAAATCTATAATGTAATTGTCACGGCTCATGCTTTTGnAATGATTTTCTTCATAGTAATACCTATTATAATTGGANGTTTTGGAAATTGATTAATTCCNTTGATATTAGGGGCTCCTGATATAGCTTTCNCTCGTATAAACAATATAAGATTTTGGNTGNTTCCCCCNTCTTTAACNTTACTCCTTANCAGANGATTANTTGAAANAGGAGTAGGAACGGGATGAACTGTTTACCCTCCTTTGGCNTCNNNAATTGCCCATNNTGGAGCTTCTGTAGATTTAGGTATCTTTTNTCTTCATCTTGCTGGGGCTTCATCAATTTTAGGGGNAGTAAACTTTATTACNACTNTNATTAATATACGNGCNTCCGGAATTACTATGGATCGTGTGCCTTTATTTGTTTGNTCTGTTTTANTAACCGCGGTTCTTCTTCTCTTATCTTTACCTGNT
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Culicidae was found to be postive for Wolbachia.
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