An improved PCR-based method for site directed mutagenesis using megaprimers
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An improved PCR-based method for site directed mutagenesis using megaprimers. / Brøns-Poulsen, J; Petersen, N E; Hørder, M; Kristiansen, K.
In: Molecular and Cellular Probes, Vol. 12, No. 6, 1998, p. 345-8.Research output: Contribution to journal › Journal article › peer-review
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TY - JOUR
T1 - An improved PCR-based method for site directed mutagenesis using megaprimers
AU - Brøns-Poulsen, J
AU - Petersen, N E
AU - Hørder, M
AU - Kristiansen, K
N1 - Keywords: DNA Primers; DNA-Directed DNA Polymerase; Humans; Hydroxymethylbilane Synthase; Mutagenesis, Site-Directed; Point Mutation; Polymerase Chain Reaction
PY - 1998
Y1 - 1998
N2 - An improved protocol for site-directed mutagenesis based on the two-step polymerase chain reaction (PCR) megaprimer method is described. Compared to previously published protocols, the protocol described in this article ensures consistently a success rate of at least 85% with essentially no introduction of unwanted secondary mutations. The essential features of this protocol include an optimization of the template-primer amounts and ratio that allows the use of a reduced number of PCR cycles and the use of proof-reading thermostable DNA polymerases.
AB - An improved protocol for site-directed mutagenesis based on the two-step polymerase chain reaction (PCR) megaprimer method is described. Compared to previously published protocols, the protocol described in this article ensures consistently a success rate of at least 85% with essentially no introduction of unwanted secondary mutations. The essential features of this protocol include an optimization of the template-primer amounts and ratio that allows the use of a reduced number of PCR cycles and the use of proof-reading thermostable DNA polymerases.
U2 - 10.1006/mcpr.1998.0187
DO - 10.1006/mcpr.1998.0187
M3 - Journal article
C2 - 9843651
VL - 12
SP - 345
EP - 348
JO - Molecular and Cellular Probes
JF - Molecular and Cellular Probes
SN - 0890-8508
IS - 6
ER -
ID: 11254093