Amplicon-seq of AsCas12f_edit_3
Genomic DNA was extracted 72 hrs post-transfection using a Ezup Column Animal Genomic DNA Purification Kit (Sangon Biotech). Regions surrounding the target loci were amplified using a Phanta Max Super-Fidelity DNA Polymerase Mastermix (Vazyme). PCR products were gel purified and re-annealed. The indels were detected by using T7 endonuclease I (Vazyme) digestion. For amplicon sequencing, the library was generated by a two-step amplification method. The PCR products were purified by 1.5VAHTS DNA Clean Beads (Vazyme) to remove primer dimers and subjected to Illumina HiSeq sequencing (PE150).