amplicon sequencing of sgTRAC-site-2_ABE8e edited HEK293T cells
Multiplex PCR was conducted using Phanta polymerase (Vazyme) in a 25 l reaction volume, comprising genomic DNA and 6.8 l of a primer mixture (10 primer pairs, each at 2.5 M). Following 25 amplification cycles, PCR products were resolved on a 1.5% TAE agarose gel and extracted using the E.Z.N.A. Gel Extraction Kit (Omega Bio-Tek). The amplicons were subsequently pooled and ligated to sequencing adapters using the VAHTS Universal DNA Library Prep Kit (Vazyme), adhering to the manufacturer's instructions. A final PCR amplification of 10 cycles was performed according to the kit's protocol.