Cas9 efficiency for translocation induction
The genomic DNA extracted from a pool of 100 primary transformants was used for analysis via NGS. To test for Cas9 efficiency, primers with 6 bp overhangs were designed to amplify over the nuclease cutting sites using the Q5 High-Fidelity DNA polymerase (New England Biolabs, NEB). The amplicons were purified using the peqGOLD Cycle-Pure Kit (Peqlab), pooled and sent for sequencing via the Illumina HiSeq platform at GATC Biotech.