Figure 2
Quantitative real-time PCR validation and results for NEO copy number determination, using MGB-assay (in red) and SYBR-assay (in green). (A) Twenty-two ES clones (abscissa, numbers from 1 to 22) showed a nNEO value between 0.8 and 1.2 (ordinate). One genomic DNA from a homozygous knockout mouse was used as control (CTRL 2NEO, nNEO = 2.2). Error bars indicate triplicates delta-delta Ct standard deviations. (B) Real-time PCR results: 45 ES clones are plotted on the graph, for each assay using a log2 scale. The dotted line indicates a nNEO threshold of 1.2, the upper cutoff for considering the presence of a single NEO copy. Samples tested with either of the two assays had similar nNEO values, as shown by the similar population plots.