Green Fruitworm

May 29, 2026

Sample information
Picture
Entry by: Pranav A.
Location
Collection date 04/22/2026
Captive / Cultivated? Wild-caught
Group Thomas Jefferson High School for Science and Technology
Observations

Larval-stage green fruitworm.
Putative identification Arthropoda Insecta Lepidoptera Noctuidae Orthosia Orthosia hibisci

Methods
Extraction kit Qiagen DNeasy Blood and Tissue Kit
DNA extraction location Whole arthropod
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniPCR
Buffer TBE
DNA stain SYBR Safe
Gel images
Protocol notes

Protocol:

Your teacher has already prepped for you:        

  • 1x TBE buffer
  • Gels with 26 wells
  • A microtube of loading dye

 

Student safety and prep:

  1. Be sure to wear gloves, goggles, and your lab coat. Tie your hair back if needed.
  2. Make sure the PCR tubes you have are YOURS and they are for CO1/arthropod (labeled with a C!)

 

Load your gel:

  1. Your teacher will load your ladder.
  2. Pipette 1.4 uL droplets onto parafilm loading dye (one 1.4 uL droplet per sample–7-8 drops for most groups).
  3. Pick up 7 uL of your first CO1 sample, pipet up and down on the drop of loading dye to mix, then load the samples into each well of the top row as follows:
Lane Sample Check when complete Notes
1 LEAVE BLANK
2 ___ ___ 1C
3 ___ ___ 2C
4 ___ ___ 3C
5 ___ ___ 4C
6 Ladder Your teacher adds this
7 ___ ___ 5C Positive control Drosophila
8 ___ ___ 6C Negative control Drosophila
9 ___ ___ 7C Positive control DNA
10 ___ ___ 8C Water
11-13 LEAVE BLANK

 

  1. As your teammates finish loading the top row, use your second strip of Parafilm to create another eight 1.4 uL droplets of loading dye.
  2. Clean your space, pick up your Wolbachia samples and return your CO1 samples

 

  1. Repeat step 6 with Wolbachia PCR samples:
Lane Sample Check when complete Notes
1 LEAVE BLANK
2 ___ ___ 1W
3 ___ ___ 2W
4 ___ ___ 3W
5 ___ ___ 4W
6 Ladder Your teacher adds this
7 ___ ___ 5W Positive control Drosophila
8 ___ ___ 6W Negative control Drosophila
9 ___ ___ 7W Positive control DNA
10 ___ ___ 8W Water
11-13 LEAVE BLANK

 

  1. Place the orange cover over the electrophoresis system. Match positive and negative electrodes. Be sure the lid sits flush with the base.
  2. Press the “Run” button. Run is on the right side of the device. The light bulb is the blue light button that you should activate when you check your results.
  3. Conduct electrophoresis for 15-25 minutes. Check progress every 10 minutes or so.
  4. See Schoology for ladder details.

Results
Wolbachia presence No
Confidence level High
Explanation of confidence level

The band of the caterpillar pretty much exactly matched the COI band of the controls with a clear band, but the Wolbachia trial did not have any bands at all for the caterpillar while the positive control did have clear bands.
Wolbachia 16S sequence
Arthropod COI sequence Download FASTA    Download AB1
Summary The Orthosia hibisci was found to be negative for Wolbachia.
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