Three pairs of oligonucleotide primers specific to hptII, Cas9-eGFP, and AtFT were used in PCR to determine the presence of T-DNA integration in genomic DNA. Reactions contained 100 ng of genomic DNA, 0.5 μM forward primer, 0.5 μM reverse primer, 200 μM deoxynucleotide triphosphates (dNTPs), sterile distilled water (GibcoBRL), 1× DreamTaq buffer, and 1.5 U of DreamTaq (Thermo Fisher Scientific). Products were resolved on a 1% tris-acetate-EDTA agarose electrophoresis gel containing ethidium bromide and visualized using an ultraviolet transilluminator.

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