Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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Saturation Mutagenesis by Codon Cassette Insertion

(Protocol summary only for purposes of this preview site)

Saturation mutagenesis by cassette insertion introduces a library of site-specific changes into a specific DNA sequence within a target gene and is especially useful for analyzing the effect of specific residues on the structure and function of a protein. In general, cassette insertion mutagenesis involves excising a target region by restriction enzyme cleavage at two flanking sites and inserting a custom oligonucleotide containing the desired mutation in place of the excised fragment. This approach is effective for generating a single substitution at a single site, but it is not practical if the goal is to generate a library of mutants containing all possible amino acid substitutions at a single site because of the costs involved in synthesizing a complete canon of oligonucleotides.


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(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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