Bridge_HEK293T_editing
Genomic DNA from samples were isolated by the DNeasy Blood & Tissue Kit (Quiagen) following the manufacturer's protocol. Locus-specific primers were used to generate targeted amplicons for deep sequencing. First, input genomic DNA was amplified using the AccuPrime Pfx DNA Polymerase (Thermo Fisher Scientific, 12344024). The first PCR employed primers containing Illumina adapter overhangs. A second PCR was carried out to introduce sample-specific p5p7 barcodes. PCR products were purified using the NucleoSpin Gel and PCR Clean-up kit (Macherey-Nagel) following the manufacturer's protocol. The final library pool was quantified using a Qubit 3.0 fluorometer (Invitrogen), and sequencing was performed on an Illumina MiSeq platform (150 bp, paired-end reads).