Will cloning efficiency increase if I use a longer incubation time for the In-Fusion Cloning reaction?
Will cloning efficiency increase if I use a longer incubation time for the In-Fusion Cloning reaction?
No, an increase in the In?Fusion reaction time is not recommended. It may generate uneven single-stranded regions at the ends of the cloning insert and vector, resulting in inefficient annealing of the homologous overlaps, thus reducing cloning efficiency.