TY - JOUR
T1 - PCR amplification of up to 35-kb DNA with high fidelity and high yield from λ bacteriophage templates
AU - Barnes, Wayne M.
PY - 1994/3/15
Y1 - 1994/3/15
N2 - 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 λ DNA template.
AB - 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 λ DNA template.
UR - http://www.scopus.com/inward/record.url?scp=0028211009&partnerID=8YFLogxK
U2 - 10.1073/pnas.91.6.2216
DO - 10.1073/pnas.91.6.2216
M3 - Article
C2 - 8134376
AN - SCOPUS:0028211009
VL - 91
SP - 2216
EP - 2220
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 6
ER -