The mitochondrial genome (mtDNA) is a circular, intron-free, double-stranded, haploid DNA strand 16.5 kb in length that encodes 37 genes. mtDNA copy number (mtDNA-CN) is a biomarker for mitochondrial dysfunction associated with overall mortality and a number of age-related diseases, including cardiovascular disease, chronic kidney disease, and cancer. The importance of mtDNA-CN measurement to disease suggests the potential for mtDNA-CN to be a useful biomarker in the clinic.

The RayBio^® Human Absolute Mitochondrial DNA Copy Number Quantification Kit (H-mtDNA-CNQ) is a Taqman™ probe-based qPCR assay for the specific quantitative measurement of human mtDNA-CN. Both the primer and probe of mtDNA specifically target one of the most conserved regions on human mtDNA, with no amplification of any off-target sequence on nuclear genomic DNA. Meanwhile, both primer and probe of the novel single copy gene (IFNB1) specifically recognizes and amplifies a 110 bp-long nuclear genomic DNA and serves as a reference for data normalization. The optimized PCR formulation provides superior specificity and efficiency with a wide linear dynamic range. This kit allows for the determination of human mitochondrial DNA copy number in one reaction, by the comparison of mitochondrial (mt) and nuclear (n) DNA using various genomic DNA from cultured cells, tissue, saliva, scraped cells, urine, spinal fluid, lavage fluid, blood, etc.

• No off-target amplification on nuclear DNA
• Superior specificity and efficiency
• Wide linear dynamic range
• Measures mtDNA in one reaction
• Compatible with various human samples

• Fluorescence qPCR instrument capable of reading FAM (494nm maximum absorption, 518nm maximum emission) and VIC (532nm maximum absorption, 552nm maximum excitation) channels
• Sterile nuclease-free pipette tips (barrier tips recommended)
• Microfuge tubes
• Compatible PCR Plate
• Microcentrifuge
• Vortex Mixer

Contact our technical support team for questions about compatibility: techsupport@raybiotech.com

1. Prepare all reagents, samples and standards as instructed.
2. Load the PCR reaction mix and samples.
3. Seal the PCR reaction tubes or plate.
4. Set up the PCR Amplification.
5. Export the results.
6. Calculate mtDNA copy number.

The kit can be stored at -20°C for a period of 12 months prior to opening. After opening, the reagents are valid for at least 6 months if stored at -20°C. Avoid repeated freeze-thaw cycles.