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. Mice have become the most ubiquitous model organism used to study human disease. Mice are strikingly similar to humans at the genomic level and the pathophysiology of disease in mice is similar to that of humans. Moreover, mice are a cost-effective and efficient tool to enhance research and drug testing.

The RayBio^® Mouse Mitochondrial DNA Damage Quantification Kit (MTM-DQ) is a Taqman™ probe-based qPCR assay for the specific quantitative measurement of mouse mitochondrial DNA damage. This kit targets the long mtDNA region (D-Loop gene), which is the most susceptible to damage. Gene-specific Taqman™ 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)
• Microfuge tubes
• Compatible PCR plate or PCR tubes
• 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 non-mouse and non-amplification control samples have higher Cq values than mouse DNA or are undetermined, which means there is high specificity for mtDNA amplification.

CV < 5% for Cq value and CV < 15% for mtDNA damage when the input DNA ≥ 0.3 ngreaction.

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.