ZFIN Image: VanLeuven et al., 2018, Figure 1.

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Figure Caption

Figure 1. Workflow overview of PCR and PAGE for screening CRISPR-Cas9-induced mutations in zebrafish.

F0-injected zebrafish are grown to adulthood and outcrossed to a wild-type zebrafish. 12 embryos from this outcross are sacrificed for genomic DNA extraction and PCR analysis of the region encompassing the target site. PCR products are directly run on a 15% polyacrylamide gel. This gel represents an outcross in which the F0-injected founder is carrying a single 10-bp deletion at gad2 exon 1 that is transmittable at a frequency of ∼33% to the F1 generation. The second pair of bands that are noted with an asterisk are heteroduplexes. These heteroduplexes are seen with all heterozygous samples for all alleles and are not a reflection of nonspecific primer binding.

Acknowledgments

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