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. 1994 Mar 15;91(6):2216-20.
doi: 10.1073/pnas.91.6.2216.

PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates

W M Barnes  1
Affiliations

PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates

W M Barnes. Proc Natl Acad Sci U S A. .

Abstract

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.

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