Figure 2.

Representative genotyping result using PCR/ribonucleoprotein (RNP) complex to select potential homozygous lines. Independent edited plants in the T₂ generation targeting AT1G72350 were analyzed using PCR/RNP complex, a mixture of in vitro synthesized sgRNA, Cas9 enzyme, and PCR products flanking target site. PCR amplicons identical to the sgRNA were completely digested by Cas9 (lanes 4, 5, 6, 9, and 11), while amplicons of homozygous or biallelic-mutants (red arrows) were not digested. Partially digested amplicons showing both cut- and uncut-bands indicated heterozygous mutants (lanes 2, 7, 8, 10, and 13).