|Captive / Cultivated?||Wild-caught|
|Group||St. John High School USD 350|
I found these Spotted Cucumber beetles on sunflowers. Before I preserved them in 70% ethanol they were a bright yellow and they blended in with the flowers really well. There were also Soldier beetles on the same sunflowers and blended in as well.
|Putative identification||Hexapoda Insecta Coleoptera|
|Extraction kit||DNeasy (Qiagen)|
|DNA extraction location||Abdomen|
|Single or Duplex PCR|
|Gel electrophoresis system||MiniPCR|
The beetle was stored in 90% ethanol for about a month before extraction.
DNA Extraction: The DNA was extracted from the abdomen. All of the Dneasy (Qiagen) protocols were followed. The extraction took several days to complete.
PCR: Mini One Taq polymerase was used.
Gel Electropheresis: All of the lanes worked except for the DNA ladder. When pipetting, the well was missed and had to be done again which might have been the reason.
|Explanation of confidence level||
All of the controls worked and were very clear. The DNA extraction protocols were followed exactly.
|Wolbachia 16S sequence||
|Arthropod COI sequence||
|Summary||The Coleoptera was found to be negative for Wolbachia.|