Table 2. Species-specific primers targeting the mitochondrial cytochrome oxidase subunit I (COI) gene in the nine target species of turtle, showing fragment length (in base pairs; bp); annealing temperature for PCR (TA) and the biomass of turtle relative to water in each tested sample, and whether eDNA was detected through PCR or qPCR (Y = yes for all replicates; N = no;— = not tested).
| eDNA detected? | |||||||
|---|---|---|---|---|---|---|---|
| Species | Primer | Sequence (5’-3’) | bp | TA | Target biomass in sample (g/L) | PCR | qPCR |
| Emydoidea blandingii | CO1-EBl-01-F | ATCATAATCTTCTTCATAGTC | 216 | 56 | 12.50 | Y | — |
| CO1-EBl-01-R | AGTTTCCAGCTAGTGGTGGA | ||||||
| Clemmys guttata | CO1-CGu-01-F | GCCATTAATAATCGGGGCACCG | 227 | 65 | 133.35 | Y | Y |
| CO1-CGu-01-R | GAATTGAAGATACACCAGCCAAA | ||||||
| Glyptemys insculpta | CO1-GIn-02-F | GCCAGTCATAATCGGTGGA | 155 | 62 | 1.88 | Y | — |
| CO1-GIn-02-R | CTGCTCCTGCTTCAACCCCT | ||||||
| Chrysemys picta | CO1-CPi-01-F | GAAATTGACTCGTACCAATG | 230 | 60 | 3.18 | Y | — |
| CO1-CPi-01-R | CACCCCTGCTAAGTGGAGAG | ||||||
| Graptemys geographica | CO1-GGe-01-F | GTTATTATTGCTCTTAGCATC | 202 | 56 | 375.00 | Y | — |
| CO1-GGe-01-R | GGCTGGAGATTTTATGTTAA | ||||||
| Sternotherus odoratus | CO1-SOd-01-F | CGCCTGAGCAGGCATAATTG | 299 | 65 | 0.65 | Y | — |
| CO1-SOd-01-R | CTGCACCTGCTTCAATTCCA | ||||||
| Chelydra serpentina | CO1-CSe-01-F | TGTTATAATTGGGGGCTTTGGA | 197 | 60 | 248.00 | Y | Y |
| CO1-CSe-01-R | GAGCTATGTTTCCAGATAGT | 10.71 | Y | N | |||
| Apalone spinifera | CO1-ASp-02-F | CATCTGGCCGGAGTATCGTC | 231 | 65 | 150.00 | Y | — |
| CO1-ASp-02-R | GTCTCCACCTCCTGAGGGAT | ||||||
| Trachemys scripta | CO1-TSc-01-F | GGGAACTGACTCGTGCCATTA | 178 | 65 | ~ 3.75 | Y | — |
| CO1-TSc-01-R | GGGCTAAATTTCCGGCTAAT | ||||||