Cottrell S F
Biochim Biophys Acta. 1981 Oct 27;655(3):390-5. doi: 10.1016/0005-2787(81)90050-2.
A method is described for estimating rapidly the relative proportion of total DNA that is of mitochondrial origin in small quantities of the yeast, Saccharomyces cerevisiae. This procedure involves the mechanical disruption of cells followed by the addition of small amounts of radioactively labeled yeast nuclear and mitochondrial DNA to the lysate. Both labeled and unlabeled DNAs are then co-extracted from the mixture and separated into nuclear and mitochondrial DNA components by poly(L-lysine) Kieselguhr column chromatography. The resulting specific radioactivities of each species of DNA, when compared to the amount of labeled DNA initially added, or related to the relative proportion of unlabeled nuclear and mitochondrial DNA in the original cell sample. The isotope dilution procedure reported here is shown to be both reproducible and to reflect the true relative concentration of each species of DNA in this yeast.
本文描述了一种快速估算少量酿酒酵母线粒体DNA在总DNA中相对比例的方法。该方法包括机械破碎细胞，然后向裂解液中加入少量放射性标记的酵母核DNA和线粒体DNA。接着将标记和未标记的DNA从混合物中共提取出来，并通过聚（L-赖氨酸）硅藻土柱色谱法分离成核DNA和线粒体DNA组分。将得到的每种DNA的比放射性与最初加入的标记DNA量进行比较，或者与原始细胞样品中未标记核DNA和线粒体DNA的相对比例相关联。本文报道的同位素稀释法具有可重复性，并且能够反映该酵母中每种DNA的真实相对浓度。
