Takara Human Mitochondrial DNA (mtDNA) Monitoring Primer Set

The Human Mitochondrial DNA (mtDNA) Monitoring Primer Set is designed to quantify the relative number of copies of human mitochondrial DNA (mtDNA) by real-time PCR, using nuclear DNA (nDNA) content as a standard. 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.

The Human Mitochondrial DNA (mtDNA) Monitoring Primer Set is designed to quantify the relative number of copies of human mitochondrial DNA (mtDNA) by real-time PCR, using nuclear DNA (nDNA) content as a standard. 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 induced pluripotent stem (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 iPS cells 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 Terra qPCR Direct TB Green Premix or TB Green Premix Ex Taq II (Tli RNaseH Plus).

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Overview

• Includes primer sets for detection of two mtDNA targets (ND1, ND5) and two nDNA targets (SLCO2B1, SERPINA1)
• Specific to human mtDNA; primer sets will not amplify pseudogenes
• Designed for use with Terra qPCR Direct TB Green Premix or TB Green Premix Ex Taq II (Tli RNaseH Plus)

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Applications

• Monitoring differentiation of hepatocytes induced from iPS cell lines

• Quantifying the ratio of human mtDNA to human nuclear DNA

Components

Storage

–20°C. Use within 1 year.

If using product repeatedly within the span of 1 month, store at 4°C. However, take care to avoid contamination as this product does not contain preservatives.

Data analysis

To aid data analysis, please download and use the mtDNA Copy Number Calculation Tool. This excel file can also be accessed through the product table: Details > Documents > User Manuals.

References

Yu, Y. et al. Hepatocyte-like cells differentiated from human induced pluripotent stem cells: Relevance to cellular therapies. Stem Cell Res. 9, 196–207 (2012).

Additional product information

Please see the product`s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.