Münzner P, Voigt J
Institut für Allgemeine Botanik und Botanischer Garten, Universität Hamburg, F.R.G.
J Biochem Biophys Methods. 1989 May;18(3):183-93. doi: 10.1016/0165-022x(89)90003-1.
High concentrations of KI were found to efficiently protect RNA against degradation by RNases. When a sufficient amount of solid KI was added to cell lysates or subcellular fractions (9 g per 10 ml), the solutions could be stored at room temperature for several days without measurable degradation of mRNA. Ribonucleic acids were selectively sedimented when these KI-containing solutions were centrifuged at 72,000 x g for 24 h. The RNA pellets were found to be readily dissolved in bidistilled water and the redissolved RNA could be immediately submitted to oligo(dT)-cellulose chromatography to isolate the poly(A)-containing RNA. However, extraction with phenol/chloroform was found to be necessary, if total RNA or poly(A)-minus RNA was to be analysed. This procedure was found to be superior to other methods currently in use - especially with respect to the isolation of intact, translatable high-molecular-weight mRNA.
已发现高浓度的碘化钾（KI）能有效保护RNA不被核糖核酸酶降解。当向细胞裂解液或亚细胞组分中加入足量的固体KI（每10毫升加入9克）时，溶液可在室温下保存数天，mRNA无明显降解。当这些含KI的溶液在72,000×g离心24小时时，核糖核酸会被选择性沉淀。发现RNA沉淀很容易溶解于双蒸水中，重新溶解的RNA可立即用于寡聚（dT）-纤维素柱层析以分离含聚腺苷酸（poly(A)）的RNA。然而，如果要分析总RNA或不含poly(A)的RNA，则发现有必要用苯酚/氯仿进行提取。已发现该方法优于目前使用的其他方法——尤其是在完整的、可翻译的高分子量mRNA的分离方面。
