Do DyNAzyme™ I, II and EXT DNA Polymerases, and Phusion® Polymerase add the non-template dependent 3-A overhang?
DyNAzyme™ DNA Polymerases are capable of adding a 3′-overhang. However, the efficiency at which the extra base is added is sequence-dependent. The extra base is usually A, however, the enzyme can also incorporate other bases or even more than a single base at a sequence-spesific manner. To enhance TA-cloning it may be helpful to include an additional incubation period of 10-30 minutes at 72 °C to the end of cycle protocol.
