B-4 Emma Field Cricket

October 30, 2024

Sample information
Picture
Photos by: Lucas S
Location
Collection date 09/24/2024
Captive / Cultivated? Wild-caught
Group Edmund Burke School
Observations

The Cricket has a yellow body with a shiny shell with marks. It has long, thin antennae, large back legs, and segmented legs and body. The eyes are prominent, and its snout has small appendages. The cricket was caught in a semi-urban park with lush pastures where the bug resides contrasting heavily with the loud urban backdrop. The season was fall and it was caught at night with a temp of about 72 degrees Fahrenheit.

 
Putative identification Arthropoda Hexapoda Insecta Orthoptera Rhaphidophoridae Diestrammena Diestrammena asynamora

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

 

DNA Extraction

  1. The DNA extraction went well for the most part. The only issues of note are that we added about 50ul of extra Buffer to B-2, B-3, and B-5. We also spilled a little bit of B-5.

PCR”:

  1. There was one notable mistake during the PCR, we forgot to switch pipette tips between samples possibly contaminating them.

Gel electrophoresis, Arthropod CO1

  1. The ladder was marked on all Gels
  2. Samples B-1 through B-6 were positive for arthropod DNA 
  3. The positive and negative controls were also marked positive and water was marked negative

 

Gel electrophoresis, Wolbachia

  1. B-1 through B-5 were negative and B-6 was positive
  2. The controls worked with both negative and water controls marked negative and the positive control marked positive
  3. There were no notable mistakes or errors

Results
Wolbachia presence No
Confidence level Medium
Explanation of confidence level

There was a moment when we forgot to switch pipettes between samples but it doesn’t seem to have affected the experiment. The controls worked as planned further proving the validity of our gels.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Diestrammena asynamora was found to be negative for Wolbachia.
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