The maintenance of mitochondrial genomic integrity is a prerequisite for proper mitochondrial function. Excessive oxidative stress can cause mitochondrial DNA (mtDNA) to be damaged by loss of its supercoiled structure. Accumulation of lesions in mtDNA is believed to be one of the causes of energy crisis in aging tissues as well as with numerous diseases including neurodegenerative disorders, cardiovascular diseases and cancer. Damage to mtDNA is also a meaningful biomarker for evaluating genotoxicity of drugs and environmental toxins. 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 Mitochondrial DNA Damage Quantification Kit (MTR-DQ) is a Taqman™ probe-based qPCR assay for the specific quantitative measurement of rat mitochondrial DNA damage. This kit targets the long mtDNA region (D-Loop gene), which is the most susceptible to damage. Gene-specific TaqmanTM probe qPCR is highly sensitive because of the use of “long” PCR methodology that permits the quantitative amplification of fragments of genomic DNA. As a result, very low levels of lesions can be detected, permitting the study of mtDNA damage using genomic DNA from cultured cells, tissue, saliva, urine, blood, etc.

• Detects low levels of mtDNA lesions
• Quantitative amplification of genomic DNA fragments
• Suitable for various sample types

• Fluorescence PCR instrument capable of reading FAM channel (494 nm maximum absorption, 518 nm maximum emission) and JUN channel (606 nm maximum absorption, 618 nm maximum emission)
• Sterile nuclease-free pipette tips (barrier tips recommended) and microfuge tubes
• Compatible PCR Plate
• Microcentrifuge
• Vortex Mixer
• Pipettes

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.