Citizen salmon is still here and progressing with LAMP. We had talked about LAMP a bunch in the past, but are finally making progress despite some rough starts.
We had chosen to move forward with LAMP because its isothermal (so light on the equipment needs) and suppose to result in visible-turbidity if positive (no equipment necessary for analysis).
Unfortunately with the salmon primers we had designed we weren’t getting any visible turbidity after 1-2 hours of incubation. We would leave our samples incubating overnight, then add SYBRsafe to the tubes to see if any DNA amplified. This is what we got. All positives and negatives growing like crazy!
We think that our primers are self-amplifying, which really brings our primer-designing ability into question. So what next?
Well, lets test our primer design skills with an easier target. At SoundBio, we have a circular piece of DNA called pPSU1 that can be cut to various sizes to form a DNA ladder. We thought lets cut the circular DNA once to make it linear and practice primer design the pPSU1 DNA.
David designed primers using PrimerExplorer and here are the results:
Hurray!!! Lanes going left to right:
- LMW DNA Ladder
- pPSU1 LAMP with DNA, no incubation
- pPSU1 LAMP with DNA, 15 mins
- pPSU1 LAMP with DNA, 30 min
- pPSU1 LMAP no DNA, 15 mins
Finally, controls that work!!!
That’s all for now. Thanks everyone!