NGS data showing editing efficiency in protoplasts
Target regions were amplified through two rounds of PCR. The first PCR employed site-specific primers, with the products (typically 10-fold diluted) serving as templates for the second round. The second PCR utilized primers containing unique 6-bp barcodes at their 5 termini to generate relatively short amplicons (170250 bp). Products from each sample were pooled in near-equimolar amounts for library preparation. Sequencing was performed commercially (GENEWIZ, Suzhou, China) on an Illumina Novaseq PE150 platform.