The Human Mitochondrial DNA (mtDNA) Monitoring Primer Set is designed to quantify the relative number of copies of human mitochondrial DNA (mtDNA) using nuclear DNA (nDNA) content as a standard by real-time PCR. This kit contains primer pairs for the amplification of four regions: two primer pairs for detecting mtDNA and two primer pairs for detecting nDNA. These primers will not amplify pseudogenes. As the primers are human-specific and will not amplify DNA from mice, human mtDNA content can be monitored in iPS cells or other cells cultured with feeder cells of mouse origin. This allows the Human Mitochondrial DNA (mtDNA) Monitoring Primer Set to be used in drug discovery applications where induced pluripotent stem cells (iPSCs) are used as a source of differentiated liver cells, since hepatocytes have a higher ratio of mtDNA to nuclear DNA than pluripotent cells.
Using the four primer pairs in this set for real-time PCR, relative quantification of mtDNA is determined from the difference in Ct values for mtDNA and nDNA. This product provides a simple method for monitoring changes in the mtDNA copy number as mtDNA has been reported to increase during differentiation of pluripotent cells, such as iPS cells, into hepatic cells. The primer pairs in this set are compatible with our qPCR master mixes.