GRP9 Anisodactylus Binotatus

May 26, 2025

Sample information
Picture
Photos by: Brayden N.
Location
Collection date 04/30/2025
Captive / Cultivated? Wild-caught
Group California Academy of Mathematics and Science
Observations

When caught, the beetle tried to dig out of its container.
Putative identification Arthropoda Insecta Coleoptera Carabidae Anisodactylus Anisodactylus binotatus

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Abdomen
Single or Duplex PCR Duplex Reaction
Gel electrophoresis system Standard electrophoresis system Bio-Rad
Buffer 1X TAE
DNA stain UView
Gel images
Protocol notes

Gel 1
Lane
1 – PCR MW Ladder (failed)
2 – centipede (lamyctes emarginatus)
3 – pillbug (armadillidium vulgare)
4 – woodlouse spider (dysdera crocata)
5 – ground beetle (anisodactylus binotatus)
6 – Arthropod Positive Control
7 – Arthropod Negative Control
8 – Drosophila Positive Control
9 – Drosophila Negative Control

Gel 2
Lane
3 – PCR MW Ruler
6 – MiniOne Ladder

Our first gel’s ladder did not run successfully, so we ran another gel with the same ladder and and another MiniOne ladder and compared the relative mobilities of the two gels to determine the base pairs of the first gel.

Results
Wolbachia presence No
Confidence level Medium
Explanation of confidence level

Although our controls worked,  we are unsure of why our ladder did not run, so the results of our gel are somewhat questionable. However, all of the samples ran correctly, with aligned bands that were only in expected locations
Wolbachia 16S sequence
Arthropod COI sequence Download FASTA   
NNNTNNNNTNTTTGNAGCATGATCAGGAATAGTAGGGACTTCTTTAAGCATATTAATTCGAGCTGAATTAGGAACTCCTG GTGCATTAATTGGTGACGATCAGATTTATAATGTTATTGTTACTGCTCATGCTTTTGTAATAATTTTTTTTATAGTAATA CCTATTATAATTGGAGGATTTGGAAATTGATTAGTACCTTTAATATTAGGTGCTCCTGATATAGCATTTCCTCGAATAAA TAATATAAGTTTTTGATTATTACCTCCATCATTAACCCTTCTTTTAATGAGAAGAATGGTTGAAAGAGGAACTGGGACTG GTTGAACAGTTTACCCTCCTTTATCATCTGGTATTGCCCATGGAGGAGCTTCAGTAGATTTAGCTATTTTTAGATTACAT TTAGCTGGAGTATCTTCAATTTTAGGGGCAGTAAATTTTATTACAACAATTATTAATATACGATCAGTAGGTATAACATT TGATCGTATACCATTATTTGTTTGATCAGTAGGAATTACTGCATTACTTTTATTATTATCTTTACCAGTACTAGCAGGAG CAATTACTATATTATTAACAGATCGAAATTTAAATACTTCTTTTTTTTGATCCTGCTGGAGGAGGAGACCCAATTTTATA CCAACATTTATTTTGATTTTTTGGTCACCNGNNAAGTTNNAA
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Anisodactylus binotatus was found to be negative for Wolbachia.
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