51 Life Technologies
™
| Genome editing
locus-specific double-strand breaks
Figure 13. GeneArt
®
genomic cleavage detection assay. To detect either an indel or a mutation within a specific
sequence of DNA, the region is first amplified using primers specific for that region. A second nested PCR can be
performed to increase sensitivity. After heating the sample and reannealing the PCR products, amplicons containing
indels or other changes in sequence will result in the formation of heteroduplexes with amplicons containing unmodified
sequences. When these heteroduplexes are treated with an endonuclease that only cleaves in the presence of a
mismatch, two pieces of DNA of known size are generated, which can be detected by agarose gel electrophoresis.
