Experimental design for SDF and cell clone generation.

<p>A) SDF sequence is homologous to the entire wild type eGFP coding sequence. SDF-PCR-WT, 876 bp long was generated by PCR amplification with primer pair 1F/1R (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030851#pone-0030851-t001" target="_blank">Table 1</a>). SDF-DIG-WT, 752 bp long, was obtained by <i>HindIII</i> and <i>XhoI</i> digestion of pCR-2.1 vector. C/T transition, responsible of fluorescence switching off, is showed. B) Sequencing analysis showing wild type (WT; top panel) and mutated (Mut; bottom panel) pCEP4-eGFP in C1 and D1 cell clones, respectively. Arrows indicate the modified base (C→T). C) FACS density plot of C1 (WT; top) and D1 (Mut; bottom) respectively. D) pCEP4-eGFP copy number determination for each cell clone.