title: Combination primer polymerase chain reaction for multi-site mutagenesis of close proximity sites.
abstract: We describe a rapid and efficient polymerase chain reaction procedure for multi-site-directed mutagenesis for cases in which the sites to be mutated are in close proximity. The combination primer polymer chain reaction method is based on a multi-site directed mutagenesis protocol together with a splicing by overlapping extension polymerase chain reaction protocol. several different combinations of multiple mutations were successfully performed with this method and are reported in this study.
authors.affiliation: Natlmmune A/S, Copenhagen, Denmark. phj@natimmune.dk
authors.forename: Pia Hønnerup
authors.initials: PH
authors.lastname: Jensen
authors.forename: Dietmar
authors.initials: D
authors.lastname: Weilguny
mesh_terms: D000595:Amino Acid Sequence; D004247:DNA / Q000235:genetics; D017931:DNA Primers / Q000138:chemical synthesis / Q000737:chemistry; D005822:Genetic Vectors; D006801:Humans; D050606:Mannose-Binding Protein-Associated Serine Proteases / Q000737:chemistry / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D013696:Temperature; D013698:Templates, Genetic; D013997:Time Factors
chemical_list: D017931:DNA Primers; D004247:DNA; D050606:Mannose-Binding Protein-Associated Serine Proteases