Gryllinae GRP6 (3)

May 28, 2025

Sample information
Picture
Photos by: Isabella B.
Location
Collection date 05/01/2025
Captive / Cultivated? Wild-caught
Group California Academy of Mathematics and Science
Observations

Found in a backyard located in the grass. The temperature was around 64 degrees Fahrenheit.
Putative identification Arthropoda Insecta Orthoptera Gryllidae Gryllus

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Partial abdomen
Single or Duplex PCR Duplex Reaction
Gel electrophoresis system MiniOne
Buffer 1X TAE
DNA stain UView
Gel images
Protocol notes

Lane 1: Long Beach cricket 1, Lane 3: Long beach cricket 2, Lane 5: Hawthorne cricket 1, Lane 7: Hawthorne cricket 2

Results
Wolbachia presence Unknown
Confidence level High
Explanation of confidence level

One Long Beach cricket produced one band at 53 mm, which is similar to the negative control, with a band also at 53 mm. This could mean that LB cricket 2’s DNA extraction was successful and showed that it was negative for Wolbachia. Since we could produce bands for most of the controls besides the ladder, we must have made a mistake with DNA extraction. Additionally, since our ladder didn’t run, but we had bands, this must have resulted in an improper loading of the ladder. The database search did not detect a clear primer binding site, suggesting potential issues with PCR amplification. This is most likely due to a lack of appropriately lysed cells, and we should have increased the grinding time with the pestle.
Wolbachia 16S sequence
Arthropod COI sequence
Summary
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