Combination primer polymerase chain reaction for multi-site mutagenesis of close proximity sites.
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.
Jensen
Pia Hønnerup
PH
Natlmmune A/S, Copenhagen, Denmark. phj@natimmune.dk
Weilguny
Dietmar
D
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
D017931:DNA Primers; D004247:DNA; D050606:Mannose-Binding Protein-Associated Serine Proteases