Amplicon sequencing of Solanum lycopersicum: leaf tissue
The gRNA target regions were amplified using gene-specific primers with adapter sequences. Gene-specific amplicons were diluted ten-fold in H2O before they were used as template for the addition of sample-specific indexes using adapter primers with unique barcodes. Equal volumes of each indexed amplicon reaction were pooled and subsequently gel-purified using the Monarch DNA Gel Extraction Kit (New England Biolabs). A single Illumina sequencing library was prepared using the xGen DNA Library Prep MC Kit (Integrated DNA Technologies) and sequenced on the MiSeq System (Illumina).