Protocol by Myles Wagner ([email protected]) | Last updated: 7/8/2025

Pre-sampling

Once at a site with Cassiopea, record latitude and longitude, water temp, and salinity

Take a photo or take note of the environment type they are found in (in situ documentation

Collecting the Cassiopea

How you collect the Cassiopea will depend on environment type and personal sampling preference, but here is a basic protocol:

Once you find a cassiopea by either snorkeling or wading, scoop up cassiopea with your palm on their bell (preferably wearing neoprene gloves to avoid stings)
Place into bucket with seawater for holding
Getting Cassiopea above 6 cm in diameter is preferred

Place Cassiopea in shallow container with clean water
Take two photos, one of the bell, and one of the arms (Photo Guidelines below)
Measure bell diameter (I measure from bell crease to bell crease but if others have a different preference we could measure from bell edge to bell edge)
Count rhopalia number (this is sometimes used to delineate species)
Count Arm number (Also sometimes used to delineate species)
Sex (This is why the Cassiopea should be larger than 6cm)
Using dissection scissors, clip off ~250mg of arm tissue (approximately 1cm long) from the end of the arm
IMPORTANT: Dry off the tissue sample as much as possible before putting it into the tube. I use chemwipes to blot off as much salt water as possible.
- Extra salt water could mess with the preservative solution and make extractions more difficult.
Place the sampled tissue into a 2mL tube with 1500μL DNA/RNA-shield.
- DNA/RNA shield will keep the sample stable at room temperature, but if you have access to a fridge/freezer, store the samples there. If DNA/RNA shield not possible, other nucleic acid preservatives can be ethanol/RNAlater/silica.

Add Data to shared Google Sheet: (Cassiopea Global Phylogeny Sampling)

Take two photos, one of the bell, and one of the arms (example photos attached)

Include the labeled tube you will use for sampling in the photo
These photos are for morphological categorization and so should be clear enough to determine: number and size of vesicles, colors of vesicles, presence of brooding vesicles, and shape of bell markings along the rim of the bell
You may need to take a third photo from the side to get bell markings clearly
Include a reference either ruler, or color standard in the photo if possible
If you have the means to take standardized photos with consistent lighting and color cards, that is preferred but not required

Add Data to shared Google Sheet: (Cassiopea Global Phylogeny Sampling)

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In a situation where you can get standardized photos:

In the field:

*Bare minimum is snapping a photo of the top and bottom to have a sense of their morphology

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