Primers |
Profile |
Plant
rbcL primers:
(rbcLaF / rbcLa rev) |
- Initial step: 94°C 1 minute
- 35 cycles of the following profile:
- Denaturing step: 94° C 15 seconds
- Annealing step: 54° C 15 seconds
- Extending step: 72° C 30 seconds
- One final step to preserve the sample:
4° C ad infinitum
|
Plant
matK primers:
(matK-3F / matK-1R) |
- Initial step: 94°C 3 minutes
- 41 cycles of the following profile:
- Denaturing step: 94° C 30 seconds
- Annealing step: 48° C 40 seconds
- Extending step: 72° C 1 minute
- Additional extending step: 72° C 10 minutes
- One final step to preserve the sample:
10° C ad infinitum
|
Plant
Plant-ITS primers:
(nrITS2-S2F / nrITS2-S3R) |
- Initial step: 95°C 2 minutes and 30 seconds
- 35 cycles of the following profile:
- Denaturing step: 95° C 30 seconds
- Annealing step: 56° C 30 seconds
- Extending step: 72° C 30 seconds
- Additional extending step: 72° C 10 minutes
- One final step to preserve the sample:
10° C ad infinitum
|
Plant (algae-specific)
tufA primers:
(tufA_F / tufA_R) |
- Initial step: 94°C 4 minutes
- 35 cycles of the following profile:
- Denaturing step: 94° C 1 minute
- Annealing step: 54° C 30 seconds
- Extending step: 72° C 1 minute
- One final step to preserve the sample:
10° C ad infinitum
|
Vertebrate (non-fish)
Vertebrate primer cocktail:
(VF1_t1 / VF1d_t1 / VF1i_t1/
VR1d_t1 / VR1_t1 / VR1i_t1) |
- Initial step: 94°C 1 minute
- 35 cycles of the following profile:
- Denaturing step: 94° C 15 seconds
- Annealing step: 54° C 15 seconds
- Extending step: 72° C 30 seconds
- One final step to preserve the sample:
4° C ad infinitum
|
Vertebrate (fish)
Vertebrate primer cocktail:
(VF2_t1 / FishF2_t1 /
FishR2_t1 / FR1d_t1) |
- Initial step: 94°C 1 minute
- 35 cycles of the following profile:
- Denaturing step: 94° C 15 seconds
- Annealing step: 54° C 15 seconds
- Extending step: 72° C 30 seconds
- One final step to preserve the sample:
4° C ad infinitum
|
Invertebrate
COI primers:
(LCO1490 / HC02198) |
- Initial step: 94°C 1 minute
- 35 cycles of the following profile:
- Denaturing step: 95° C 30 seconds
- Annealing step: 50° C 30 seconds
- Extending step: 72° C 45 seconds
- One final step to preserve the sample:
4° C ad infinitum
|
Fungi
ITS primers:
(ITS1F / ITS4) |
- Initial step: 94° C 1 minute
- 35 cycles of the following profile:
- Denaturing step: 94° C 1 minute
- Annealing step: 55° C 1 minute
- Extending step: 72° C 2 minutes
- One final step to preserve the sample:
4° C ad infinitum
|
Fungi (lichen specific)
ITS primers:
(ITS1F_(Gad) / ITS4) |
- Initial step: 94° C 1 minute
- 35 cycles of the following profile:
- Denaturing step: 94° C 30 seconds
- Annealing step: 54° C 30 seconds
- Extending step: 72° C 45 seconds
- One final step to preserve the sample:
4° C ad infinitum
|