Progress, Problems and Primers

We've had some solid progress, and we've run into some thorny issues that we need to solve in order to complete the vision for field detectable jellyfish. That's what innovation is all about.

First, after a few false starts, we established a small colony of moon jellies in a Kreisel aquarium.

 

This gives us a reliable source of water samples known to contain Aurelia aurita DNA, and the moons that didn't survive give us a ready source of known tissue samples. Since establishing the aquarium we've been trying to create LAMP primers for A. Aurita, with no success so far. LAMP primers are notoriously difficult to get right, so we will persist trying new combinations on different parts of the mitochondrial genome.

What happened to the tuna? The published LAMP primers for Thunnus albacares turned out to generate a lot of false positives (the primers alone would amplify), so we've abandoned that approach in favor of A. aurita.

At this point we planned to be perfecting the sampling process, but to do that we need to be able to test sample amplification. Since we don't yet have LAMP primers that work, we've decided to proceed with sampling development using qPCR. Primers for qPCR are much easier to develop (2 primers instead of 4 to 6 LAMP primers). In our next lab note we should be able to report out on primer improvments and sampling methods.