Blunting Protocol for NEB PCR Cloning Kit ( E1202 )
Mix the following components in a sterile microfuge tube : Reactions containing restriction enzyme digested DNA are incubated at room temperature for 15 minutes .
Reactions with sheared / nebulized DNA or PCR products are incubated at room temperature for 30 minutes .
Immediately inactivate enzyme in the blunting reaction by heating at 70°C for 10 minutes .
Proceed directly to the ligation step using the Quick Ligation Kit ( NEB # M2200 ) or standard T4 DNA Ligase ( NEB # M0202 ) .
