FIGURE 2.

Isolation of transgene-free T₁ plants from the sbeI-c31 and sbeIIb-c1 mutant lines. (A) A schematic description of the location/direction of the primer sets used for the analysis for the presence of T-DNA sequences in T₁ plants of sbeI-c31 (B) and sbeIIb-c1 lines (C) T₀ lines may contain CRISPR/Cas9 complex in their genome. However, self-pollination of T₀ lines allows the removal of the T-DNA following segregation in the T₁ generation plants. DNA fragments of OsU3:gRNA, Cas9 and hptII were not detected in some T₁ plants of sbeI-c31 lines (Number 4, 6, 7, 13, 14, and 19) (B) and T₁ plants of sbeIIb-c1 (Number 4, 11, 17 and 18) (C), respectively. The control PCR product was amplified from the endogenous ACTIN gene, indicating that the genomic DNAs used have sufficient quality for PCR. WT, wild type DNA control.