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. Rat has become one of the most ubiquitous model organisms used to study human disease. This is partly due to the striking similarities of rats and humans at the genomic level, but also because the pathophysiology of disease in rats is similar to that of humans.

The RayBio^® Rat Absolute Mitochondrial DNA Copy Number Quantification Kit (MTR-CNQ) is a Taqman™ probe-based qPCR assay. Both the primer and probe of mtDNA specifically target one of the most conserved regions on rat mtDNA. Meanwhile, the primer and probe of the single copy gene (AT1) specifically recognizes and amplifies a 210 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 rat 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 (494 nm maximum absorption, 518 nm maximum emission) and VIC (532 nm maximum absorption, 552 nm 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.

CV < 5% for Cq value and CV < 20% for mtDNA damage when the input DNA ≥ 0.3 ng reaction.

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.