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Abstract for the GeneEditor™ in vitro Site-Directed Mutagenesis System
The GeneEditor™ in vitro Site-Directed Mutagenesis System is a
high-efficiency system for the generation and selection of oligonucleotide-directed
mutations. This system uses antibiotic selection to obtain a high frequency of
mutants. Selection Oligonucleotides provided with the GeneEditor™ System
encode mutations that alter the ampicillin resistance gene, creating a new
additional resistance to the GeneEditor™ Antibiotic Selection Mix. In the
GeneEditor™ System protocol, the Selection Oligonucleotide is annealed to a
single- or double-stranded DNA template at the same time as a mutagenic
oligonucleotide. Subsequent synthesis and ligation of the mutant strand links
the two oligonucleotides. The resistance to the GeneEditor™ Antibiotic
Selection Mix encoded by this mutant DNA strand facilitates selection of the
desired mutation. The system will work with any cloning vector that contains
ampicillin resistance as a selectable marker. Mutants generated using this
system retain ampicillin resistance and gain resistance to the GeneEditor™
Antibiotic Selection Mix.
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