Analysis of off-target coDel. (A) A schematic of the coDel experiment targeting the TWI2 locus. A part (1022 bp) of the TWI2 locus was cloned into pMcoDel and introduced into the new MACs of conjugating cells at 7 hpm. Deletions at the endogenous TWI2 locus in the MAC were analyzed by genomic PCR. Deletions at the TWI6 and TWI7 loci, which share 90.6 and 65.5% identities at the nucleotide level with the TWI2 locus, respectively, were also analyzed by genomic PCR. (B–D) Results of genomic PCR detecting deletions at the endogenous TWI2 (B), TWI6 (C), and TWI7 (D) loci in the MAC. PCR products from the same progeny lines were separated in the same lane positions. The number of progeny lines showing any deletions at the endogenous target loci was determined. The solid arrowheads indicate PCR products from the target loci without deletions. The open arrowheads show the expected positions of PCR products from the target loci with deletions corresponding to the exact target sequences.