Asiopus minimus 2B1

April 2, 2025

Sample information
Picture
Photos by: Aryan G
Location
Collection date 02/27/2025
Captive / Cultivated? Wild-caught
Group Thomas Jefferson High School for Science and Technology
Observations

The arthropod is a black beetle, no other coloring. It is small, width at 0.5 inches. Beetle is slightly curved, legs are still intact. There are ridges among the abdomen.
Putative identification Arthropoda Insecta Coleoptera

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

Removed exoskeleton and separated abdomen

Used sand to homogenize

Did not use RNAse

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

Confidence level of absence of Wolbachia is high – no band of DNA corresponding to Wolbachia. Controls all worked as expected.

Quality score of 56 (CO1), Continuous Read Length of 656 (CO1)

Blast confirmed Asiopus minimus was the species
Wolbachia 16S sequence Download FASTA    Download AB1
TGATCCGGAATAGTAGGAACGTCACTAAGACTTTTAATTCGAGCTGAACTTGGCAACCCTGGATCTTTAATTGGAGACGATCAAATTTATAATGTAATCGTCACAGCTCACGCTTTTATCATAATTTTCTTCATAGTTATACCCATTATAATTGGAGGATTTGGAAATTGATTAGTTCCACTAATGCTAGGAGCCCCTGATATAGCATTCCCTCGAATAAATAACATAAGATTTTGATTATTACCACCATCCCTTACTCTTTTACTAATAAGAAGAATTGTAGAAAATGGAGCTGGTACAGGATGAACAGTTTACCCCCCACTGTCATCTAACATCGCCCATGGTGGATCATCTGTAGATCTGGCAATTTTTAGACTACATTTAGCAGGAATTTCTTCTATTTTAGGTGCAGTTAACTTCATTACAACAGTTATTAATATACGACCACAAGGAATAACATTTGACCGAATACCTCTATTCGTTTGAGCAGTAGTAATTACTGCAGTATTGCTTCTCCTATCTCTTCCAGTACTAGCAGGTGCAATTACTATACTATTAACAGATCGAAACATTAATACCTCATTCTTCGATCCAGCAGGTGGAGGAGATCCAATCCTTTATCAACATTTATTCTGATTTTTTGGTCACCCC
BLAST at The Wolbachia Project   BLAST at NCBI
Arthropod COI sequence
Summary The Coleoptera was found to be negative for Wolbachia.
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