Objective:

PRJ003-Aiptasia-STB’s primary objective is to monitor and establish patterns for the variability acrross four strains (CC7-Endo, CC7-SSB01, CC7-Apo, and H2-Endo)

Sampling Time Points

The Stability experiment will track the community in each strain over time with each timepoint serving as a doubling from the previous time point. New Tanks with n=40 anemones for each strain were started on 12/03/21. Anemones were left to acclimate for two days at which point samples were taken at:

• T=0 days (12/6/21)
• T=1
• T=2
• T=4
• T=8
• T=16
• T=32 days

# Sampling Protocol 5 anemones per time point per condition were placed in MP biomedicals tubes containing lysing matirces B(0.4g) and D(0.8g) and 500ul of DNA/RNA Shield (Zymo). Samples were then bead beat at 8m/s for two cyces of 60s with a 5 min pause between cycles.

3 Seawater samples were also taken at each sampling using 0.2 micron filter. for each sample 1L of ASW wash pushed through the filter using a sterile syringe; filters were then sealed at either end using parafil and frozen at -80 until downstream processing

for the first week of the experiment anemones were not fed so as not to have the presence of artemia influence the results. Following, Aiptasia were fed 2x weekly and Artemia samples were taken at time of feeding. 100ul of artemia was sampled and proccessed in the same prepared bead beating tubes under the same condition.