Ligation Protocol for NEB PCR Cloning Kit ( E1202 )
Mix the first 3 components of the reaction .
Add Cloning Master Mix ( 2X ) , 5 μl , to a total of 10 μl per ligation reaction .
Incubate at room temperature ( 25°C ) for 5–15 minutes .
Incubate on ice for 2 minutes .
Transform immediately into NEB 10 - beta Competent E . coli or store at - 20°C .
