PAM depletion assay of SpaCas12f1
100 ng of the PAM library plasmid were electroporated into 50 L of electrocompetent cells. Colonies were harvested and plasmids were extracted using MolPure Plasmid Mini Kit (Yeasen). A two-step amplification method was used to generate the amplicon sequencing library, and the PCR products were purified by using VAHTS DNA Clean Beads (Vazyme Biotech) for Illumina HiSeq sequencing.