Extended Data Fig. 7: Identification and analysis of the inversion for wheat mutants of Fragment 20-22.
a, Representative agarose gel of PCR products for genotyping of the 7.4-kb inversion. The 752-bp band and the 606-bp band are the desired bands for the left and right junction of the inversion, respectively, while the 598-bp and the 760-bp bands are the expected size for the wild-type sequence. WT, wild-type. T0-1, T0-2, T0-3, T0-4, T0-5, T0-7, T0-8, T0-9, T0-10, and T0-11 were homozygous. b, Representative Sanger sequencing chromatograms of the 7.4-kb inversion mediated by DualPE in wheat plants. T0-2 was identified as the precise inversion mutant at both Junction-L and Junction-R. In contrast, T0-5 contained a single nucleotide polymorphism (SNP) at the intended Junction-R, likely resulting from byproducts associated with the pegRNA scaffold. Meanwhile, T0-28 exhibited insertions and deletions (indels) at the targeted Junction-L, which can be attributed to the presence of micro-homologous sequences. c, Representative agarose gel of PCR products for genotyping of the 19.2-kb inversion. The 325-bp band and the 730-bp band are the desired bands for the left and right junction of the inversion, respectively, while the 695-bp and the 360-bp bands are the expected size for the wild-type sequence. WT, wild-type. T0-7 is homozygous. d, Representative Sanger sequencing chromatograms of the 19.2-kb inversion mediated by DualPE in wheat plants. T0-7 was identified as the precise inversion mutant at both Junction-L and Junction-R. In contrast, T0-10 contained a single nucleotide polymorphism (SNP) at the intended Junction-L, likely resulting from byproducts associated with the pegRNA scaffold. e, Representative Sanger sequencing chromatograms of the precise 82.8-kb inversion mediated by DualPE in wheat plants (T0-42).
