Designing and Generating CRISPR-Cas Mutants: Difference between revisions
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[[ Category: Cloning ]] | |||
[[ Category: Molecular Biology ]] | |||
[[ Category: Genome Editing ]] | |||
[[ Category: CRISPR-Cas ]] | |||
==Overview== | ==Overview== | ||
We are using the double-nickase mutation strategy. This involves the generation of two CRISPR-Cas plasmids per mutation, each of which nicks a different strand, spaced apart by about 40 nt. To generate a point mutant, you can also include a piece of DNA containing the mutation and allow for homologous recombination. | We are using the double-nickase mutation strategy. This involves the generation of two CRISPR-Cas plasmids per mutation, each of which nicks a different strand, spaced apart by about 40 nt. To generate a point mutant, you can also include a piece of DNA containing the mutation and allow for homologous recombination. | ||