Tribolium Castaneum (Red Flour Beetle)

November 21, 2023

Sample information
Picture
Photos by: Jacey C
Location
Collection date 09/07/2023
Captive / Cultivated? Wild-caught
Group Hawaii Community College
Observations

Three red flour beetles trapped and collected in local office in Kailua-Kona, Hawaii using bug zapper. Used whole arthropod for PCR.
Putative identification Arthropoda Hexapoda Insecta Coleoptera Tenebrionidae Tribolium Tribolium castaneum

Methods
Extraction kit Edwards Buffer
DNA extraction location Whole arthropod
Single or Duplex PCR Single Reaction
Gel electrophoresis system Edvotek Gel Electrophoresis
Buffer TBE
DNA stain SYBR Safe
Gel images
Protocol notes

Gel Information:

Well #1: DNA ladder
Well #2: Red Flour Beetle 1
Well #3: Red Flour Beetle 2
Well #4: Red Flour Beetle 3
Well #5: Drosophila (positive Wolbachia Control)
Well #6 Drosophila (negative control)
Well #7: DNA positive control
Well #8 Denucleased water (negative DNA control)

Top half of gel used Arthropod CO1 gene primer. Only the first red flour beetle correctly amplified the arthropod CO1 gene. Bottom half of gel used Wolbachia 16s primer. None of the red flour beetles had a Wolbachia gene amplified.

Extracted DNA from three red flour beetles, ran CO1 PCR and Wolbachia 16sRNA PCR on each sample. One tested positive for CO1, did not test positive for Wolbachia. None of the other arthropods tested positive for CO1 or 16sRNA.

Results
Wolbachia presence No
Confidence level Low
Explanation of confidence level

Only 1/3 of the beetles correctly amplified an arthropod CO1 gene. DNA was very low quality when sequenced. Not confident in the arthropod results, denucleased water control appears contaminated with arthropod DNA.
Wolbachia 16S sequence
Arthropod COI sequence Download AB1
Summary The Tribolium castaneum was found to be negative for Wolbachia.
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