Additional file 3: Figure S3. of Aneuploidy screening of embryonic stem cell clones by metaphase karyotyping and droplet digital polymerase chain reaction

Use of standard real time PCR for chromosome counting. The figure shows copy number of Chr 8 measured by qPCR with various input quantities of genomic DNA as template. The experiment demonstrates that although there is linearity across the range of concentrations relevant to the DNA preparations assayed, the assay is not sufficiently robust for the screen. Error bar amplitude varies with gDNA input and standardizing input is challenging due to the disparity of growth rates between ES cell clones. qPCR assays were performed in triplicates. SEM are represented by error bars. Both literature and our own experience concur in concluding that standard qPCR does not allow for sufficient accuracy to reliably identify clones worthy of microinjection. (PDF 25 kb)