The parameters of the curve all seem acceptable, typically reactions results are above 0.9852, -3.1 to -3.6 slope, and 90-110% reaction efficiency. The results seem to be what I would expect from eDNA and the dilution series seems to be working well. But the lack of a plateau is making me doubt the validity of the results. Do you think this is an issue. What could cause it?

I am using primers designed for another project, temperatures that multiple authors have tested with them and confirmed, and reagents otherwise identical to their experiments. Increasing the number of cycles has no effect, and I have tried experimenting with reagent concentrations and total reaction volume without any luck. I am considering changing mastermix next, which will be pricey...

Thanks for the help,

Sam