Wild Type reads
Multiplex PCR-based amplicon sequencing was performed to assess on-target genome-editing efficiency in tomato (Solanum lycopersicum cv. M82) hairy roots. Genomic DNA was extracted from Agrobacterium rhizogenestransformed hairy roots carrying CRISPRCas9 genome-editing constructs or from WT control roots. Amplicons spanning approximately 500 bp upstream and downstream of sgRNA target sites were amplified using high-fidelity PCR, pooled at equimolar concentrations, barcoded, and sequenced using PacBio SMRT long-read sequencing. Barcodes 16 correspond to CRISPR-edited biological replicates, and barcode 7 corresponds to WT control.