MR. HANCOCK -- Well, I don't need the extension time, Your Honor.
Subsequently, the optimal annealing temperature, annealing time, extension time and cycle numbers were evaluated.
The total number of cycles was 40 with the final extension time of 10 min.
The annealing temperature, extension time, and primer concentration of the traditional PCR were optimized.
To control the false positive signals, the hybridization condition, DNA polymerase, extension time and primers design were optimized.
An extension time of 1 min. was used in the PCR reactions.
We used extension time which was sufficient to cover the full-length of the plasmid.
The long extension time (10 min) was necessary due to the hairpin structure of the template.
The extension time, y (min), was set at 1 min per 1 kb.
Touchdown from 65°C to 52°C, followed by 9 cycles at 52°C; extension time of 4 minutes.
The reaction conditions may need to be adjusted if other polymerases are used, especially the extension time.
