HomeFAQsVent DNA Polymerase and Deep Vent DNA Polymerase produce blunt DNA ends, but my cloning depends on the primer extension product having an adenine single-base 3´ overhang.
FAQ: Vent DNA Polymerase and Deep Vent DNA Polymerase produce blunt DNA ends, but my cloning depends on the primer extension product having an adenine single-base 3´ overhang.
Treat your blunt end products with Taq DNA Polymerase (NEB #M0267, NEB #M0273). After primer extension with Vent DNA Polymerase, phenol:chloroform extract and precipitate the DNA with 2 volumes of isopropanol. Resuspend in 1X ThermoPol Reaction Buffer, add 1 unit of Taq DNA Polymerase and 200 µM dATP; mix well and incubate at 72°C for 20 minutes.
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