Department of Biological Sciences, Mississippi State University, Starkville, MS 39762, USA.
Viruses. 2019 Apr 3;11(4):324. doi: 10.3390/v11040324.
Plant-viroid interactions represent a valuable model for delineating structure-function relationships of noncoding RNAs. For various functional studies, it is desirable to minimize sample variations by using DNA, RNA, and proteins co-purified from the same samples. Currently, most of the co-purification protocols rely on TRI Reagent (Trizol as a common representative) and require protein precipitation and dissolving steps, which render difficulties in experimental handling and high-throughput analyses. Here, we established a simple and robust method to minimize the precipitation steps and yield ready-to-use RNA and protein in solutions. This method can be applied to samples in small quantities, such as protoplasts. Given the ease and the robustness of this new method, it will have broad applications in virology and other disciplines in molecular biology.
植物类病毒相互作用为阐明非编码 RNA 的结构-功能关系提供了有价值的模型。对于各种功能研究，通过从相同样品中共同纯化的 DNA、RNA 和蛋白质来最小化样品变化是很理想的。目前，大多数共纯化方案依赖于 TRI 试剂（Trizol 作为常见代表），并且需要进行蛋白质沉淀和溶解步骤，这使得实验处理和高通量分析变得困难。在这里，我们建立了一种简单而强大的方法来最小化沉淀步骤，并在溶液中获得即用型 RNA 和蛋白质。该方法可应用于小量样品，如原生质体。鉴于这种新方法的简便性和稳健性，它将在病毒学和分子生物学的其他领域有广泛的应用。
