Tanaka K, Yazawa K, Najima T
Faculty of Pharmaceutical Science, University of Tokyo, Japan.
Biomed Chromatogr. 1989 Nov;3(6):246-50. doi: 10.1002/bmc.1130030604.
A simple and precise method was developed for the separation of nucleosides including modified nucleosides and oligonucleotides. Nineteen kinds of nucleosides were completely separated by HPLC using an ODS column (TSK-gel ODS 80TM) and aqueous mobile phases. The RNA molecule was digested by base restrictive RNase (RNase A, RNase T1) and the digests were separated chromatographically into each oligonucleotide. The nucleoside composition of an oligonucleotide was then determined by this analytical system. It is thus possible to fit the oligonucleotide in the original RNA molecule by using modified bases as markers. The reaction site of quinacrine mustard for tRNA(Phe) (from yeast) could be determined by this analytical system.
开发了一种简单而精确的方法用于分离包括修饰核苷和寡核苷酸在内的核苷。使用ODS柱（TSK-gel ODS 80TM）和水性流动相，通过高效液相色谱法（HPLC）将19种核苷完全分离。RNA分子用碱基限制性核糖核酸酶（核糖核酸酶A、核糖核酸酶T1）消化，消化产物通过色谱法分离成各个寡核苷酸。然后通过该分析系统测定寡核苷酸的核苷组成。因此，通过使用修饰碱基作为标记，可以将寡核苷酸与原始RNA分子进行匹配。通过该分析系统可以确定喹吖因氮芥对（来自酵母的）tRNA（Phe）的反应位点。
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