Pilliod_et_al_2013_MER_DryadData_7-31-13

We handled, stored, and extracted eDNA from 0.45 nm Cellulose Nitrate filter paper the same way for each experiment using protocols described in Pilliod et al. (2013). We extracted DNA from half of each filter paper using the Qiashredder/DNeasy Blood & Tissue DNA extraction kit method described in Goldberg et al. (2011). We used the quantitative PCR assay described in Pilliod et al. (2013) to estimate the amount of eDNA in each sample. We used the QuantiTect Multiplex PCR Mix (Qiagen, Inc., Gaithersburg, Maryland, USA) with recommended multiplexing concentrations and parameters on an Applied Biosystems 7500 Fast Real-Time PCR System to conduct the assay. We used 2 µL of DNA extract in each reaction and ran all reactions in triplicate. Standard curves were constructed from whole genomic DNA extracted from tail tissue and diluted to 0.001, 0.01, and 0.1 ng. Average r2 for curves used in this study was 0.99. We considered a test result negative if there was no exponential phase at any point during 50 cycles. See Pilliod et al. (2013) for additional methodological details.