title: UMI_HEK4_Dup262
design: First, a UMIs appending linear amplification reaction was performed using 300 ng genomic DNA and 0.04 M of single forward primer in a reaction volume of 50 l. The reaction conditions were: 95  for 3 min, 5 cycles of 65  for 1 min and 72  for 2 min, 1 cycle of 95  for 15 s, and then another 5 cycles of 65  for 1 min and 72  for 2 min. This reaction was cleaned up using 1.5  beads (Vazyme), and 5 ng purified DNA was subject to PCR with 0.04 M of forward and reverse primers in a final reaction volume of 10 l. The forward primer anneals to the upstream of the UMI sequence and is not specific to the genomic loci. We programmed the PCR reaction to be: 95  for 3 min, 28 cycles of 95  for 15 s, 65  for 10 s, and 72  for 45 s. Then 2 l UMI-appended PCR products was subject to another standard PCR with 0.04 M of forward and reverse primers in a final reaction volume of 50 l. Primers were used to append indexes and adaptors for Illumina Platform (Novaseq 6000). We calculated the efficiency as the number of edit products UMIs out of the number of wt products UMIs.