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线粒体DNA拷贝数的定量分析:分析前因素

Quantification of mitochondrial DNA copy number: pre-analytical factors.

作者信息

Andreu Antonio L, Martinez Ramiro, Marti Ramon, García-Arumí Elena

机构信息

Unitat de Patologia Mitocondrial, Centre d'Investigacions en Bioquímica i Biologia Molecular, Institut de Recerca Hospital Universitari Vall d'Hebron, Barcelona, Spain.

出版信息

Mitochondrion. 2009 Jul;9(4):242-6. doi: 10.1016/j.mito.2009.02.006. Epub 2009 Mar 9.

Abstract

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.

摘要

线粒体DNA(mtDNA)含量对于理解许多细胞过程至关重要。从样本采集到DNA提取的几个分析前因素会影响mtDNA拷贝数的测量。在本研究中,全血样本产生的mtDNA拷贝数高于血沉棕黄层样本。mtDNA含量受所用细胞分离方法以及采血与细胞分离之间的时间影响。因此,必须使用相同类型的样本建立参考值。至于DNA分离和纯化方法,手工苯酚法可能会随机给出错误的高值。QIAamp DNA Mini试剂盒提供了最具可重复性的mtDNA/nDNA产量。

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