Cricket

October 23, 2023

Sample information
Picture
Location
Collection date 09/12/2023
Captive / Cultivated? Captive / Cultivated
Group Palm Beach Atlantic University
Observations

This insect was bought at a pet store. It was held captive in a cage with others of its kind. Owners of the store tried to mimic the insect’s natural habitat as best as possible in a container.
Putative identification Arthropoda Hexapoda Insecta Orthoptera Gryllidae

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

For the gel electrophoresis a 1kb ladder was used. 4uL of ladder was loaded along with 10uL of each sample combining corresponding tubes of Wolbachia and arthropod primers. The gel ran for 45 minutes. Lane 1- ladder, Lane 2- ant, Lane 3- Cricket Lane 4 (+) control, Lane 5  (-) control, Lane 6 – (+) DNA and Lane 7- water

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

Analysis: My control worked but my cricket sample did not. There were bands at approximately 575bp indicative of arthropods in all the lanes except those of the negative controls. Then bands at 316bp indicative of the Wolbachia bacteria was only seen in the two positive controls and the ant sample. This indicates that the insect chosen (cricket) had its DNA amplified but that DNA was not infected by Wolbachia.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Gryllidae was found to be negative for Wolbachia.
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