It is critical to design the genome editing strategy to facilitate genotyping of clones, since this step can otherwise be costly and rate-limiting. We have written this protocol using the most generic case, in which the genome editing strategy does not incorporate specific features that facilitate identification of the desired modification. As a result, Steps 37–47 use the Surveyor nuclease to determine if efficient modification occurred in pooled genomic DNA. Sanger sequencing is used in Steps 51–54 to genotype individual clones.
