Figure 4

Colony PCR results for evaluation of gene disruption. Agarose gel showing an example of colony PCR to detect mutagenesis in ets1-TALEN injected Xenopus tropicalis embryos. The bands amplified by Primer 1/3 in upper panel (~200 bp) indicate the colonies harbor the TALEN targeted sites. PCR assay was also carried out with Primer 2/3 in lower panel. The colonies that can give upper bands but fail to give lower bands were the positive colonies carrying potential mutants. The colonies giving both upper and lower bands did not carry mutants (arrows indicated). Mutations were further confirmed by sequencing.