High-pressure liquid chromatography in polynucleotide synthesis.
High-pressure liquid chromatography in polynucleotide synthesis.
复制标题
多核苷酸合成中的高压液相色谱。
作者：
H. Fritz;R. Belagaje;E. Brown;R. Fritz;R. Jones;R. G. Lees;H. Khorana
反相高压液相色谱（HPLC）使用含有具有键合的十八烷基的微粒材料的柱，已经被开发为用于分离核苷、核苷酸，并且特别是被保护的寡核苷酸的快速和有效的方法，所述被保护的寡核苷酸是脱氧核糖多核苷酸的逐步合成中的标准中间体。报告是广泛的研究的影响，不同的嘌呤和嘧啶碱基，保护基团，磷酸基团，和寡核苷酸的链长对它们的保留在这样的列。此外，已经描述了HPLC在逐步合成寡核苷酸d（G-G-A-A-G-C-T-T-A-A-C）中的应用。本文所述的方法适用于制备规模的分离，并显著减少（高达50%）合成寡核苷酸所需的时间。
Reverse phase high-pressure liquid chromatography (HPLC) using columns containing microparticulate materials with bonded octadecyl groups has been developed as a rapid and efficient method for the separation of nucleosides, nucleotides, and, in particular, of protected oligonucleotides which are standard intermediates in the stepwise synthesis of deoxyribopolynucleotides. Reported are extensive studies of the influence of the different purine and pyrimidine bases, of protecting groups, of the phosphate groups, and of the chain lengths of oligonucleotides on their retention on such columns. Further, the application of HPLC in the stepwise synthesis of an oligonucleotide, d(G-G-A-A-G-C-T-T-A-A-C), has been described. The methods, which are herein described, lend themselves to separations on a preparative scale and effect a marked reduction (up to 50%) in the time required for the synthesis of oligonucleotides.