Table 2.
List of primers used in the present study (without including the Illumina adapters67) and thermocycling conditions followed during the first PCRs for eDNA metabarcoding library preparation for each primer set.
| Target region | Primer (5ʹ–3ʹ) | Thermocycling conditions | References |
|---|---|---|---|
| Croc_COI | COIa: AGTATAAGCGTCTGGGTAGTC | Croc_COI: 94 °C (120 s) followed by 35 cycles at 94 °C (45 s), 48 °C (45 s), 72 °C (90 s); and a final extension of 72 °C (10 min) | 36,37 |
| COIf: CCTGCAGGAGGAGGAGAYCC | |||
| Croc_CR | drL15459: AGGAAAGCGCTGGCCTTGTAA | Croc_CR: 94 °C (120 s) followed by 33 cycles at 94 °C (30 s), 58 °C (30 s), 72 °C (45 s); and a final extension of 72 °C (7 min) | 56 |
| CR2HA: GGGGCCACTAAAAACTGGGGGGA | |||
| MiFish (12 S) | F: CCGGTAAAACTCGTGCCAGC | 12S: 95 °C (180 s), 35 cycles of 98 °C (20 s), 65 °C (15 s), 72 °C (15 s), and a final extension of 72 °C (5 min) | 25 |
| R: CATAGTGGGGTATCTAATCCCAGTTTG | |||
| VertCOI | BloodmealF1_t1: ACCACWATTATTAAYATAAARCCMC | VertCOI: 94 °C (120 s) followed by 60 cycles at 94 °C (40 s), 56 °C (40 s), 72 °C (30 s); and a final extension of 72 °C (5 min) | 61 |
| BloodmealF2_t1: ACTACAGCAATTAACATAAAACCMC | 61 | ||
| VR1_t1:TAGACTTCTGGGTGGCCAAAGAATCA | 60 | ||
| VR1d_t1:TAGACTTCTGGGTGGCCRAARAAYCA | 60 | ||
| VR1i_t1:TAGACTTCTGGGTGNCCNAANAANCA | 60 |