GRP15 Oleander Aphid

May 24, 2024

Sample information
Picture
Photos by: Maegan A., Faith D., Kristine F., Christine M.
Location
Collection date 05/15/2024
Captive / Cultivated? Wild-caught
Group California Academy of Mathematics and Science
Observations

Arthropod was found in the school garden in clusters on the stems of milkweed in front of classroom.
Putative identification Arthropoda Hexapoda Insecta Hemiptera Aphididae

Methods
Extraction kit tweezers
DNA extraction location Whole arthropod
Single or Duplex PCR Single Reaction
Gel electrophoresis system Standard electrophoresis system
Buffer TAE
DNA stain Fast Blast
Gel images
Protocol notes

Both gels ran for 35 minutes at 135 volts and 70 amps. Results were viewed w/ a white light box.

*Wells starting new ruler (Gel 1: Arthropod)

Lane 1: DNA Ladder

Lane 2: Aphids DNA

Lane 3: Ants DINA

Lane 4: (+) Arthropod Control

Lane 5: (-) Arthropod Control

Lane 6: DNA Control

Lane 7: Water

 

*Wells starting new ruler (Gel 2: Wolbachia)

Lane 1: DNA Ladder

Lane 2: Aphids DNA

Lane 3: Ants DINA

Lane 4: (+) Wolbachia Control

Lane 5: (-) Wolbachia Control

Lane 6: DNA Control

Lane 7: Water

Results
Wolbachia presence Unknown
Confidence level High
Explanation of confidence level

Both gels did not detect visual evidence of bands when running gel electrophoresis. After consideration of the protocol, the error must have occurred in the PCR process for the DNA controls and ladder were not apparent in their designated lanes. 
Wolbachia 16S sequence
Arthropod COI sequence
Summary
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