title: Ecoli_ABES4
design: On- and off-target sites were amplified from the genome. PCR amplification was performed with Phanta Max Super-Fidelity DNA polymerase (Vazyme) using primers listed in Supplementary Table X. 50 l PCR reactions were purified with TIANquick Maxi Purification Kit (TIANGEN). The PCR products were added with Illumina-compatible adapters and indices.  Final amplicon libraries were sequenced with 150-bp paired-end reads on an Illumina MiSeq instrument, and the RAW data was analyzed by the CRISPRosso2. PCR primers about 250bp upstream and downstream of on-target sites were designed and synthesized. On-target genomic regions of each site were amplified by PCR and then were analyzed with Sanger sequencing. The results were quantified using EditR