Human Absolute Mitochondrial DNA Copy Number Quantification qPCR Assay Kit

Product Description

• 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

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. 

Specificity 
The non-human and non-amplification control samples have higher Cq values than human DNA, which means there is high specificity for mtDNA and IFNB1 amplification. 

Reproducibility
CV<5% for Cq value and CV<15% for Copies/Cell when the input DNA ≥ 0.3 ng / reaction.

Performance Data

Fig.1. Standard curves of mtDNA-FAM and IFNB1-VIC amplification for different DNA concentrations (From 30 pg - 100,000 pg/reaction) of Hela cells detected by this kit. 

Fig.2. Amplification curves of mtDNA-FAM and IFNB1-VIC amplification for different DNA concentrations (From 30 pg – 100,000 pg/reaction) of Hela cells detected by this kit.