Table 2. Conditions of PCR for Eco-223, Eco-455 and tetA primers.
|
Steps |
Temperature and duration for Eco-223 and Eco-455 primer |
Temperature and duration for tetA primer |
Cycle |
|
Initial denaturation |
95°C for 5 minutes |
95°C for 5 minutes |
1 cycle |
|
Denaturation |
95°C for 1 minute |
94°C for 1 minute |
30 cycles |
|
Annealing |
56°C for 40 seconds |
57°C for 30 seconds |
|
|
Extension |
72°C for 1 minute |
72°C for 1 minute |
|
|
Final extension |
72°C for 5 minutes |
72°C for 5 minutes |
1 cycle |
|
Holding |
4°C until use |
4°C until use |
- |
Single cycle for Initial denaturation; 30 cycles for Denaturation, Annealing, Extension; Single cycle for Final extension