Chunk #25 — INTRODUCTION — Experimental design — Functional testing — Detection of indels or HDR by sequencing

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Genome engineering using the CRISPR-Cas9 system.

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Targeted genome modifications can also be detected by either Sanger or deep sequencing. For the former, genomic DNA from the modified region can be amplified with either SURVEYOR or HDR primers. Amplicons should be subcloned into a plasmid such as pUC19 for transformation, and individual colonies should be sequenced to reveal the clonal genotype.