title: Two-stage PCR protocol allowing introduction of multiple mutations, deletions and insertions using QuikChange Site-Directed Mutagenesis.
abstract: We developed a two-stage procedure, based on the QuikChange Site-Directed Mutagenesis Protocol, that significantly expands its application to a variety of gene modification experiments. A pre-PCR, single-primer extension stage before the standard protocol allows the efficient introduction of not only point mutation but also multiple mutations and deletions and insertions to a sequence of interest.
authors.affiliation: Schering-Plough Research Institute, Kenilworth, NJ 07033, USA.
authors.forename: W
authors.initials: W
authors.lastname: Wang
authors.forename: B A
authors.initials: BA
authors.lastname: Malcolm
mesh_terms: D000668:Ampicillin Resistance / Q000235:genetics; D001483:Base Sequence; D001709:Biotechnology; D017931:DNA Primers / Q000235:genetics; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000235:genetics; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D017384:Sequence Deletion
chemical_list: D017931:DNA Primers