PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates.
WM Barnes - Proceedings of the National Academy of …, 1994 - National Acad Sciences
Proceedings of the National Academy of Sciences, 1994•National Acad Sciences
A target length limitation to PCR amplification of DNA has been identified and addressed.
Concomitantly, the base-pair fidelity, the ability to use PCR products as primers, and the
maximum yield of target fragment were increased. These improvements were achieved by
the combination of a high level of an exonuclease-free, N-terminal deletion mutant of Taq
DNA polymerase, Klentaq1, with a very low level of a thermostable DNA polymerase
exhibiting a 3'-exonuclease activity (Pfu, Vent, or Deep Vent). At least 35 kb can be amplified …
Concomitantly, the base-pair fidelity, the ability to use PCR products as primers, and the
maximum yield of target fragment were increased. These improvements were achieved by
the combination of a high level of an exonuclease-free, N-terminal deletion mutant of Taq
DNA polymerase, Klentaq1, with a very low level of a thermostable DNA polymerase
exhibiting a 3'-exonuclease activity (Pfu, Vent, or Deep Vent). At least 35 kb can be amplified …
A target length limitation to PCR amplification of DNA has been identified and addressed. Concomitantly, the base-pair fidelity, the ability to use PCR products as primers, and the maximum yield of target fragment were increased. These improvements were achieved by the combination of a high level of an exonuclease-free, N-terminal deletion mutant of Taq DNA polymerase, Klentaq1, with a very low level of a thermostable DNA polymerase exhibiting a 3'-exonuclease activity (Pfu, Vent, or Deep Vent). At least 35 kb can be amplified to high yields from 1 ng of lambda DNA template.
National Acad Sciences
