@article{b75a3ef00fac11de8478000ea68e967b,
title = "An improved PCR-based method for site directed mutagenesis using megaprimers",
abstract = "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.",
author = "J Br{\o}ns-Poulsen and Petersen, {N E} and M H{\o}rder and K Kristiansen",
note = "Keywords: DNA Primers; DNA-Directed DNA Polymerase; Humans; Hydroxymethylbilane Synthase; Mutagenesis, Site-Directed; Point Mutation; Polymerase Chain Reaction",
year = "1998",
doi = "10.1006/mcpr.1998.0187",
language = "English",
volume = "12",
pages = "345--8",
journal = "Molecular and Cellular Probes",
issn = "0890-8508",
publisher = "Academic Press",
number = "6",
}