72620 Sigma-AldrichNovaQUANT™ Human Mitochondrial to Nuclear DNA Ratio Kit

The NovaQUANT™ Human Mitochondrial to Nuclear DNA Ratio Kit compares the levels of nuclear to mitochondrial DNA (mtDNA) in a human DNA sample.

Real-Time PCR provides a platform for a quantitative assay measuring and comparing mtDNA copy number to that of nuclear DNA (nDNA). This mtDNA/nDNA ratio test kit in qPCR format can be used whenever a small amount of DNA can be extracted from cells or tissues of interest. The primers in this kit have been thoroughly validated for matching amplification efficiency and for absence of amplification of mitochondrial pseudogenes.

A single assay is run using four wells, whereby each well individually tests for one of four gene targets. Resultant Cts obtained from the Real-Time instrument are used to represent the level of each gene. Nuclear gene 1 is compared to that of mitochondrial gene 1 and nuclear gene 2 to mitochondrial gene 2. With this method, ratios are calculated which can be averaged to represent the mtDNA copy number per cell.

Panel Components

• 2 ea qPCR plate with pre-aliquoted PCR primer pairs
• 100ng Human wt DNA (1 ng/µl)
• 100ng Human rho zero DNA (1 ng/µl)

This kit contains a set of four optimized PCR primer pairs targeting two nuclear and two mitochondrial genes.

The NovaQUANT Human Mitochondrial to Nuclear DNA Ratio Kit contains two qPCR plates with pre-aliquoted PCR primer pairs, Human wt DNA, and Human rho zero DNA. Please see Technical Bulletin (TB563) for other materials that are required, but not supplied, for using this kit. This kit contains enough DNA for 48 tests (using two 96-well microplates).

Storage: All materials in this kit can be kept at 4 °C for up to three months; we recommend keeping control DNAs at –20 °C for long term storage.

• Optimized qPCR protocol for sensitive, specific and linear detection of mitochondrial or nuclear target genes.

• Eliminate detection of unwarranted mitochondrial pseudogenes in your data.

• Accurately determine DNA ratios based on matched primer pair amplification efficiencies.

• Pre-aliquoted primers for automating your protocols.