Spotted cucumber beetle (Diabrotica undecimpunctata) C-5 SSP

November 21, 2024

Sample information
Picture
Photos by: Sam H.
Location
Collection date 09/17/2024
Captive / Cultivated? Wild-caught
Group Edmund Burke School
Observations

Collected Outside, 70ºF, wet ground, only one of its kind in the area. Terrestrial, urban habitiat. Yellow, spotted, small.

 
Putative identification Arthropoda Insecta Coleoptera Chrysomelidae Diabrotica Diabrotica undecimpunctata

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

Crushed, extracted abdomen, no exoskeleton. PCR used to amplify the specific genes. Dyed the samples of DNA, mixed, and put into respective gel spots.

 

Arthropod CO1: No mistakes, controls worked to perfection.

Gel electrophoresis: No mistakes, controls worked.

1-kb ladder

2- C-4

3- C-5

4- W+

5- W-

6- H2O

7 – empty

8- empty

9 – rerun of C-3

PCR: No mistake

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

Multiple tests showed no trace of Wolbachia gene, no errors made in setup or testing of this sample. Controls of W+, H2O, and W- all came out the way they were supposed to. No errors in the lead-up/process of this lab for this sample.
Wolbachia 16S sequence
Arthropod COI sequence Download FASTA   
Diabrotica undecimpunctata
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Diabrotica undecimpunctata was found to be negative for Wolbachia.
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