Article
In Vitro Mutagenesis Using Double-Stranded DNA Templates: Selection of Mutants with DpnI.
Cold Spring Harbor protocols - 1 Mar 2018
Forloni Matteo, Liu Alex Y, Wajapeyee Narendra
Abstract excerpt
In this protocol, two oligonucleotides are used to prime DNA synthesis by a high-fidelity polymerase on a denatured plasmid template. The two oligonucleotides both contain the desired mutation and have the same starting and ending positions on opposite strands of the plasmid DNA. The entire lengths of both strands of the plasmid DNA are amplified in a linear fashion during several rounds of thermal cycling,...
Topics
- DNA
- DNA Primers
- Deoxyribonucleases, Type II Site-Specific
- Mutagenesis
- Mutagenesis, Site-Directed
- Mutation
- Templates, Genetic
- Transformation, Genetic