Table 3 Moat water samples spiked with varying starting quantities of A. tomentosa DNA for environmental DNA isolation methods, estimated final concentrations, and their respective average Cq values and SD, used to calculate actual final concentrations of DNA in each sample used to assess DNA recovery from each method used
Sample starting quantity (μg) | Estimated final concentration (ng/μL) | Average Cq | Calculated final concentration (ng/μL) | Recovery (%) |
|---|---|---|---|---|
NaOAC + NaI clean up | ||||
1 | 9.9 × 10−2 | + | N/A | – |
0.5 | 5 × 10−2 | + | N/A | – |
0.25 | 2.5 × 10−2 | 13.53 ± 0.21 SD | 7.73 × 10−2 | > 100 |
0.125 | 1.25 × 10−2 | 12.49 ± 0.68 SD | 1.56 × 10−1 | > 100 |
Neg | – | ND | – | – |
Filtering – 50 mL vol | ||||
1 | 2 × 10−2 | 15.93 ± 0.18 SD | 1.53 × 10−2 | 76.7 |
0.5 | 1 × 10−2 | 17.52 ± 0.31 SD | 5.25 × 10−3 | 52.54 |
0.25 | 5 × 10−3 | 18.37 ± 0.10 SD | 2.92 × 10−3 | 58.47 |
0.125 | 2.5 × 10−3 | 19.53 ± 0.05 SD | 1.36 × 10−3 | 54.24 |
Neg | – | ND | – | – |
Filtering – 250 mL vol | ||||
1 | 8.5 × 10−3 | 16.88 ± 0.08 SD | 8.09 × 10−3 | 95.15 |
0.5 | 4.25 × 10−3 | 18.38 ± 0.11 SD | 2.94 × 10−3 | 69.25 |
0.25 | 2.13 × 10−3 | 20.35 ± 0.01 SD | 7.80 × 10−4 | 36.72 |
0.125 | 1.06 × 10−3 | 21.36 ± 0.30 SD | 3.95 × 10−4 | 37.19 |
Neg | – | ND | – | – |