Hi colleagues,

The cycle parametre for cycle sequencing of Bigdye Terminator V 3.1 cycle sequencing Kit is "96°C 1min; 96°C 10s, 50°C 5s,60°C 4min, repeat 25cycles;

60°C 4min". Some of my primers annealing temperature is 45°C - 48°C. In that case, should I change cycle paramere for cycle sequencing? For example, "96°C 1min; 96°C 10s, 45°C 5s,60°C 4min, repeat 25cycles;60°C 4min"?

Thank you in advance.