Gallagher M L, Burke W F, Orzech K
Biochem Biophys Res Commun. 1987 Apr 14;144(1):271-6. doi: 10.1016/s0006-291x(87)80506-5.
A study of the use of carrier RNA to improve precipitation of DNA from dilute solutions was conducted to define the conditions which optimize DNA recovery. Replicate samples containing labeled pBR322 and increasing concentrations of commercially-available Torula yeast RNA were ethanol precipitated at -20 degrees C for 1 h in microfuge tubes obtained from various manufacturers. Nucleic acids were pelleted by centrifugation for either 5 or 30 min, dried and resuspended. Although recovery was not identical in each type of microfuge tube, in all cases the percent recovery increased when carrier was added. In most cases, extending centrifugation to 30 min did not significantly increase recovery. Recovery of unlabeled DNA's of heterogeneous molecular weight and conformation was also enhanced by the addition of carrier RNA. DNA's recovered by this method can be successfully digested with BamHI and ligated with T4 DNA ligase.
开展了一项关于使用载体RNA改善从稀溶液中沉淀DNA的研究，以确定优化DNA回收的条件。含有标记的pBR322和浓度不断增加的市售圆酵母RNA的重复样品，在来自不同制造商的微量离心管中于-20℃用乙醇沉淀1小时。通过离心5分钟或30分钟使核酸沉淀，干燥并重悬。虽然在每种类型的微量离心管中回收率并不相同，但在所有情况下，添加载体后回收率百分比均有所增加。在大多数情况下，将离心时间延长至30分钟并不会显著提高回收率。添加载体RNA也提高了不同分子量和构象的未标记DNA的回收率。通过这种方法回收的DNA可以成功地用BamHI消化并用T4 DNA连接酶连接。
