Mitochondrial DNA (mtDNA) content quantification assay
- MtDNA and nuclear DNA extraction from samples.
- DNA quantification and quality assessment using NanoDrop spectrophotometer.
- Calculate suitable dilution factors for optimized experimental efficiency. Two different dilution factors are used to ensure accuracy.
- Set up and run the real-time qPCR reactions with mtDNA and nuclear DNA standards (no risk of amplifying pseudogenes- see figure 1).
Features of the Assay:
- Accurate results. Unique oligonucleotide primers for mtDNA and nuclear DNA are used. Samples are assayed in triplicates, at two different dilutions and compared against reference standards.
- Reliable services. Reference standard curves are run in the same experiment by using serial dilutions of pre-determined copy number of mtDNA and nuclear DNA to provide absolute quantification as well as relative quantification depending on your needs.
- Save money. No need to purchase real-time qPCR instruments or reagents or optimize conditions.
- Save time. Let our experienced technical experts do the experiments for you.
Fig.1. Duplication of the mitochondrial genome in the nuclear genome: more than 95% of mitochondrial genome is duplicated in nuclear sequences in for of pseudogenes. The mitochondrial genome sequence accession number NC-001807, was blasted against the reference sequence of the human genome using blast, the first 50 best matching sequences are shown, the top red line being an exact match to the mitochondrial genome, whereas the remaining 49 lines are regions of the nuclear genome showing a high degree of identity. Malik AN, Czajka A. Mitochondrion, 2012
- Malik AN et al. Mitochondrial DNA as a non invasive biomarker: Accurate quantification using real time quantitative PCR without co-amplification of pseudogenes and dilution bias. Biochem Biophys Res Commun. 2011 Aug 19;412(1):1-7
- Malik AN, Czajka A. Is mitochondrial DNA content a potential biomarker of mitochondrial dysfunction? Mitochondrion 2012 Oct22.,
- Malik AN, Shahni R, Iqbal MM. Increased peripheral blood mitochondrial DNA in type 2 diabetic patients with nephropathy. Diabetes Res Clin Pract. 2009;86(2):e22-4
mitoDNA®” is a registered Community Trademark of King’s College London, with number 012279981