FIG 3.
Determination of inversion efficiency by digital droplet PCR. (A) During the inversion procedure, cellular regions also undergo inversion. Primers and probes specific for regions in the beta-globin and procollagen 1 alpha genes were, respectively, designed such that they would amplify no matter the inversion state (green) or only when the 1-kb fragment was inverted (blue). (B) Duplex ddPCR was carried out on untreated (UT) and Myrcludex B-treated (+MyrB) samples (n = 3) depicted in Fig. 8, including a no-ligation negative control (No lig) and a water negative control (H2O). (C) After a threshold was defined according to water controls, absolute numbers of each noninverted (green) and inverted product (blue) were calculated. For each data point, absolute copy numbers ± 95% Poisson error are shown.