Simultaneous Quantification of Mitochondrial DNA Damage and Copy Number in Circulating Blood: A Sensitive Approach to Systemic Oxidative Stress
Figure 1
A new strategy for absolute quantification of total and damaged mtDNA. This protocol is separated into four main steps. (a) Construction of the mtDNA and nuclear DNA standards. Long fragments of genes that contained the shorter real-time PCR targets were amplified and a 10X serial dilution was made from 3 × 106 to 30 copies. (b) Preparation of the original and preheated DNA templates from lymphocytes DNA samples for the analysis of mtDNA damage and total mtDNA, respectively. (c) Real-time PCR absolute quantification analysis of mtDNA damage and total content. (d) Interpretation of the data.
