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 ).
