See post for details on extracting DNA from May 2025 FHL eelgrass + Vpec experiment.
Last spring I ran an experiment where the study question was:
Can the presence of eelgrass decrease the bacterial counts of V. pectenicida in seawater?
The treatment groups were:
| Treatment | Treatment Code | Number of bags (replicates) | Eelgrass in treatment? Yes/No |
|---|---|---|---|
| Low Dose Vibrio + Eelgrass | LDVE | 8 | Yes |
| High Dose Vibrio + Eelgrass | HDVE | 8 | Yes |
| Low Dose Vibrio | LDV | 8 | No |
| High Dose Vibrio | HDV | 8 | No |
| Eelgrass + Filtered seawater | EFSW | 8 | Yes |
samples
0.45um filters that I filtered the water from each individual replicate (n=40) through.
The filters were stored in -80C originally at FHL, but then brought down to UW last month, and have been in the FTR -80C since then.
DNA Extraction
Following similar protocols from when I worked on the Eelgrass and Eelgrass wasting disease project, I processed half of the 0.45um filter from each sample.
I followed the protocol from ZymoBIOMICS DNA MiniPrep Kit (D4300), and used the Roberts Lab Bullet Blender 5E Gold + with the 2.0mL adapters (1min at speed 12, 5 min rest, 5 times).
The DNA is eluted in 50ul of water and stored in the middle -80C in FTR.
The remaining 1/2 filters of each sample is also in the middle -80C in FTR.
Next Steps
Waiting on some supplies, but I will be processing them in mid-/ late- April to look for V. pectenicida and relative abundance across the different treatment groups.
That will help inform treatment selection for the re-iteration of this experiment this coming May at FHL, where I’ll also be adding the treatment group of shellfish alone, shellfish+eelgrass, etc., to see if shellfish can decrease bacterial counts.