Quantification of mitochondrial DNA copy number: pre-analytical factors.
Mitochondrial DNA (mtDNA) content is important for understanding many cellular processes. Several pre-analytical factors, from sample collection to DNA extraction can affect measurement of mtDNA copy number. In the present study, whole blood samples yielded a higher mtDNA copy number than buffy coat samples. mtDNA content is affected by the cell separation method used and the time between blood withdrawal and cell separation. Thus, reference values must be established with the same type of sample. As to the DNA isolation and purification method, the manual phenol method can give randomly false high values. The QIAamp DNA Mini Kit provided the most highly reproducible mtDNA/nDNA yield.
Andreu
Antonio L
AL
Unitat de Patologia Mitocondrial, Centre d'Investigacions en Bioquímica i Biologia Molecular, Institut de Recerca Hospital Universitari Vall d'Hebron, Barcelona, Spain.
Martinez
Ramiro
R
Marti
Ramon
R
García-Arumí
Elena
E
D001773:Blood Cells / Q000737:chemistry*; D004272:DNA, Mitochondrial / Q000032:analysis*; D006801:Humans; D015203:Reproducibility of Results; D013048:Specimen Handling / Q000379:methods*
D004272:DNA, Mitochondrial