Investigation and Quantification of Mitochondrial DNA Copy Number in a Single Cell by Real-Time PCR
Kodagoda T. Nimalsha Hansani
*
Department of Physiology and Human Biology, VNU University of Science, Hanoi, Vietnam.
*Author to whom correspondence should be addressed.
Abstract
The mitochondrial (mtDNA) genome is considered a small DNA molecule that consists of dozens of mitochondrial genome copies. This mitochondrial genome contains the highest mutation rate compared with the nuclear genome. Some defects are identified in the mitochondrial genomes that are caused by diseases in the aging process [1]. This higher level of mutation in the mtDNA genome determines the level of the heteroplasmic mutation in the individual cells and tissue obtained from the patients. This experiment is designed to determine the mitochondrial genome copy number using the Real-time PCR technique and indicates the specific relationship with southern blotting and competitive three-primer PCR to detect and quantify the mitochondrial DNA deletions within a single cell. This is demonstrated to investigate the patients with mitochondrial diseases and individual muscle fibers that contain the different types of abnormalities in the single and multiple deletions in the patients. This experiment is designed to investigate and quantify the deleted mtDNA in patients.
Keywords: Mitochondrial DNA, cytotoxic C-deficient cell, real-time PCR, southern blotting, three primer PCR