Why were GCN detected with conventional methods but not eDNA analysis?
If GCN are in low abundance, they may not be shedding enough DNA into the water column to enable it to be captured and amplified. Alternatively, if a pond has greater habitat complexity, then higher sampling effort may be required than what is stated in the Natural England protocol. Dilution is the prescribed strategy to resolve inhibition when identified in samples to dilute inhibitors present in the sample. However, any target DNA present will also be diluted, possibly to concentrations too low to be detected.
