TABLE 4.
Primer information and optimal conditions for Tilapia_v2_16S, R.marina_16S, and C.caroliniana_matK eDNA assays
| Assay | Gene region | Annealing (°C) | [Primer] (nM) | Amplicon (bp) |
gDNA |
sDNA |
|||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Efficiency | LOD (ng/µl) | Tm (°C ± 99.7% CI) | Efficiency | LOD (copies/µl) | Tm (°C ± 99.7% CI) | Oligonucleotide (5′–3′) | |||||
| Tilapia_v2_16S_F | 1273–1290 | 60 | 500 | 101 | 102.6%a 97.4%b | 2.65E−06a 2.81E−06b | 82.76 ± 0.69a 83.07 ± 0.52b | 96.7% | 3 | 83.02 ± 0.57 | AATGTCTTTGGTTGGGGC |
| Tilapia_v2_16S_R | 1377–1396 | TTCTGTTGCTTGGAGTTGTA | |||||||||
| R.marina_16S_F | 30–49 | 65 | 250 | 290 | 93.3% | 6.00E−05 | 80.79 ± 0.72 | 93.8% | 3 | 80.05 ± 0.41 | AGCCTGCCCAGTGACCATG |
| R.marina_16S_R | 319–338 | TGTTATGCTCCGTGGTCACC | |||||||||
| C.caroliniana_matK_F | 640–664 | 65 | 600 | 265 | 97.9% | 6.86E−04 | 76.00 ± 0.47 | 83.9% | 3 | 75.73 ± 0.60 | GCTCCTTCTTTACATCTATTGCGAT |
| C.caroliniana_matK_R | 884–903 | GGTGCCACTACAAGACGCT | |||||||||
a
O. mossambicus. bT. mariae.