Environmental DNA detection of the giant freshwater crayfish (Astacopsis gouldi)

This supplementary datasets contains amplification data for all environmental DNA samples collected as part of publication "Environmental DNA detection of the giant freshwater crayfish (Astacopsis gouldi)" submitted to "Environmental DNA" There are two separate datasets; one with the cT values of A. gouldi amplification by qPCR (amplification metadata) and the other with cT values of eDNA sample amplification during initial quality control for inhibition (Inhibition tests metadata). Each dataset corresponds to separate supplementary material documents.

Supplementary S1. Amplification of giant freshwater crayfish: All samples were tested for inhibition by dilution followed by spiked DNA tests.  All Samples were run in triplicate (including negative extraction blanks, field and negative controls) using the A. gouldi Taqman assay. We considered samples as positive if amplification crossed a common threshold determined individually within each qPCR run and if detections had CT values ≤ 55 in any of the replicates. If all technical replicates from a sample showed no amplification, then samples were amplified again. If no positive amplification was observed in the re-run qPCR, then a final qPCR was prepared with neat and diluted eDNA samples. If no amplification was observed in any of the qPCR tests then samples were deemed negative. Quantitative PCR reactions were done using the Viia™ 7 Real-Time PCR System (Applied Biosystems, Vic., Australia). 

Appendix S3. Inhibition tests of environmental DNA samples collected in this study: This supplementary dataset contains amplification data for all environmental DNA samples collected as part of this publication. All samples were tested for inhibition by dilution followed by spiked DNA tests.  All Samples were run in triplicate (including negative extraction blanks, field and negative controls) using the A. gouldi Taqman assay. We considered samples as positive if amplification crossed a common threshold determined individually within each qPCR run and if detections had CT values ≤ 55 in any of the replicates. If all technical replicates from a sample showed no amplification, then samples were amplified again. If no positive amplification was observed in the re-run qPCR, then a final qPCR was prepared with neat and diluted eDNA samples. If no amplification was observed in any of the qPCR tests then samples were deemed negative. Quantitative PCR reactions were done using the Viia™ 7 Real-Time PCR System (Applied Biosystems, Vic., Australia).