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. Author manuscript; available in PMC: 2022 Oct 19.
Published in final edited form as: Biotechniques. 2018 Jun;64(6):275–278. doi: 10.2144/btn-2018-0012

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

Figure 1.

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.