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PCR thermocycling program temperatures and time for microorganism’s species-specific primer/probe set for each gene. The optimal annealing temperature for the species-specific primers varied between primer/probe combinations.
| PCR Stage | Temp °C | Time | E. coli | Salmonella | Aeromonas | 16S |
|---|---|---|---|---|---|---|
| Enzyme Activation | 95 | 4 min | 4 min | 4 min | 4 min | 4 min |
| Denature DNA | 95 | 30 s | 30 s | 30 s | 30 s | 30 s |
| Annealing | Varies | 60 s | 60 ℃ | 60 ℃ | 62 ℃ | 56 ℃ |
