G7 – A. Vulgaris Secluded 2

May 23, 2024

Sample information
Picture
Photos by: Leoni S., Shirley J., Edgar G., Troy M., Emma Z., Jules Q.
Location
Collection date 05/13/2024
Captive / Cultivated? Wild-caught
Group California Academy of Mathematics and Science
Observations

Approx 4.5mm x 2mm x 2.5mm, exoskeleton consisting of overlapping blue-gray shells with white-green longitudinal spot pattern
Putative identification Arthropoda Crustacea Malacostraca Isopoda

Methods
Extraction kit DNeasy (Qiagen) blood and tissue kit
DNA extraction location Gonads dissected
Single or Duplex PCR Single Reaction
Gel electrophoresis system Standard electrophoresis system
Buffer TAE
DNA stain UView
Gel images
Protocol notes

Run gels at 100V constant for 45 minutes using HindIII ladder mixed with UView.

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

The arthropod DNA sequences (CO1) showed up for the insect and DNA positive samples, which not showing for the DNA negative control. This indicates that DNA extraction, PCR, and gel electrophoresis were correctly conducted without contamination, validating the procedure used for the Wolbachia 16S sequence electrophoresis. The results are further justified by the DNA positive and Wolbachia positive samples being the only samples to show a band indicating the 16S sequence on the Wolbachia PCR gel, meaning that there was no contamination specifically during the DNA extraction, PCR, and gel electrophoresis protocol concerning the Wolbachia 16S sequence.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Isopoda was found to be negative for Wolbachia.
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