Table 1.
Sample sets | Conditions | PCR result | ||
Lab. | Temperature°C | Time | COX1 | ITS |
1 | 26-28 | 3 days | ✓ | ✓ |
2 | 26-28 | 1 week | ✓ | ✓ |
3 | 26-28 | 2 weeks | ✓ | ✓ |
4 | 37 | 3 days | ✓ | ✓ |
5 | 37 | 1 week | ✓ | ✓ |
6 | 37 | 2 weeks | ✓ | ✓ |
7 | 4 | 6 months | ✓ | ✓ |
8 | -20 | 6 months | ✓ | ✓ |
Field | Conditions | COX1 | ITS | |
9 | Tropical field (28-32°C) setting for 2 weeks + transport by plane as hold luggage | ✓ | ✓ | |
10 | As in 9 and then storage at 4°C for up to 6 months | ✓ | ✓ | |
11 | As in 9 and then storage at -20°C for up to 6 months | ✓ | ✓ |
Each sample set 1-8 included 2 of each larval stage (cercariae, miracidia) and eggs preserved in 5 μl or 10 μl of RNAlater® solution. Sample sets 9-10 contain 2 miracidia collected in the field and stored in 5 μl RNAlater®. ✓ corresponds to 100% of the samples producing a positive PCR and sequences of good quality.