Rogg H, Brambilla R, Keith G, Staehelin M
Nucleic Acids Res. 1976 Jan;3(1):285-95. doi: 10.1093/nar/3.1.285.
A method is described which allows a very efficient determination of the modified nucleosides of tRNA. The technique involves enzymatic degradation of the tRNA to nucleosides at pH 7.6 and their separation by two-dimensional thin-layer chromatography on cellulose-coated aluminum foils. Based on the analysis of two mammalian tRNAs it is shown that the technique is suitable for the determination of chemically unstable nucleosides as well as the ribose-methylated compounds. At least 36 of the 45 known modified nucleosides can be separated and quantitatively determined by the method described. This procedure is especially suitable for the estimation of the nucleoside composition of unlabeled tRNAs as well as for studying the post-transcriptional modifications of tRNA.
本文描述了一种能非常高效地测定tRNA修饰核苷的方法。该技术包括在pH 7.6条件下将tRNA酶解为核苷，并通过在涂有纤维素的铝箔上进行二维薄层色谱法对其进行分离。基于对两种哺乳动物tRNA的分析表明，该技术适用于测定化学不稳定核苷以及核糖甲基化化合物。通过所述方法可以分离并定量测定已知的45种修饰核苷中的至少36种。该程序特别适用于估计未标记tRNA的核苷组成以及研究tRNA的转录后修饰。
Zotero 插件