James M, Mansbridge J, Kidson C
Anal Biochem. 1983 Jan;128(1):60-5. doi: 10.1016/0003-2697(83)90344-5.
A modification to the method for estimation of repair DNA synthesis in cultured mammalian cells using density labeling which reduces the time and labor involved is described. Centrifugation times are reduced by using high-molecular-weight DNA which is loaded onto the top of a preformed sodium iodide gradient. Cerenkov counting is used to locate the region of the gradient containing light DNA for rebanding and likewise to identify appropriate fractions from alkaline gradients for acid precipitation for scintillation counting. The procedures give results indistinguishable from cesium chloride equilibrium gradients in a shorter time, with less effort, and at a lower cost.
本文描述了一种对使用密度标记法估算培养哺乳动物细胞中修复性DNA合成的方法的改进,该改进减少了所需的时间和工作量。通过使用加载到预先形成的碘化钠梯度顶部的高分子量DNA,缩短了离心时间。用切伦科夫计数法来定位梯度中含轻DNA的区域以便重新进行区带分离,同样地,用其从碱性梯度中识别出适合酸沉淀以便进行闪烁计数的组分。这些步骤能在更短的时间内、以更少的工作量和更低的成本得到与氯化铯平衡梯度法无法区分的结果。