Ligation Protocol WITH T4 DNA Ligase ( M0202 )
Thaw the T4 DNA Ligase Buffer and resuspended at room temperature .
Set up the reaction in a microcentrifuge tube on ice .
Gently mix the reaction by pipetting up and down and microfuge briefly .
For cohesive ( sticky ) ends , incubate at 16°C overnight or room temperature for 10 minutes .
For blunt ends or single base overhangs , incubate at 16°C overnight or room temperature for 2 hours .
Heat inactivate at 65 degrees C for 10 minutes .
Chill on ice and transform 1 - 5 μl of the reaction into 50 μl competent cells .
