Fig. S6.

Screening for genome-modified flies. (Top) PCR screening after digestion with NcoI of template DNA from pools of G₁ flies derived from different G₀ (injected) individuals using a specific primer pair (ETXS/PxM) for I1056M mutation [–, nos.Cas9 DNA (negative control); +, donor plasmid template (positive control)]. (Bottom) Screening of G₂ flies from line Et15 crossed to balancer TM3Sb, PCR amplification with a generic primer pair (ETXSF/ETXSR), and digestion of 395-bp product with NcoI. The modified allele remains uncut, whereas the wild-type allele is cut in two smaller bands; positives are heterozygous at this stage.