## Objective: Sample individuals that have been treated in ABS, or ABS 2 without the priming for one week in order to determine the the susceptibility of anemones to conditions and test the if concentrations used were appropriate.

Broader Context

This will serve as preliminary data in a pre-experiment for PRJ002-Aiptasia-MM, which will allow for any needed adjustments to be made after the priming in the main experiment is complete.

Protocol

Samples total were taken at time of sampling SAM136-138 were taken from the ABS 1 plate wells A1, B2, and C1 respectively, SAM_139-141 were taken from ABS2 plate wells A3, B2, and C1 respectively. SAM_142 was taken from the stock tanks to use as a control.

All anemones were homogenized in 300ul of Filtered ASW using a a handheld 1.5ml tube pestle. 50 ul were removed from the tube and used to plate serial dilutions of homogenate on Marine Agar plates at 10⁰, 10^-1, and 10^-2 dilutions ( 50ul per plate). Samples were spread using 5 roller beads per plate and passed through a infinity motion 20 times and left in the incubator at 30C for 24 hours. the remaining 250 ul was transferred to a MP Biomedicals Lysing matrix E tube with the glass bead removed. 500ul of DNA/RNA shield was added to the lysing tubes and samples were bead beat at 8 m/s for 60 seconds twice with a five minute pause between the two cycles, then frozen at -80 in Box Auntil further processing.

Raw Data

Sample | CFU | Dilution ——-|—–|——- 136 | 2 | 10⁰ 137 | 0 | 10⁰ 138 | 1 | 10⁰ 139 | 0 | 10⁰ 140 | 0 | 10⁰ 141 | 1 | 10⁰ 142 | 51 | 10^-2