Ausserer W A, Biros M L
Dionex Corporation, Sunnyvale, CA, USA.
Biotechniques. 1995 Jul;19(1):136-9.
A strong anion-exchange high-performance liquid chromatography column, the NucleoPac PA-100, has been developed for high-resolution analysis and purification of synthetic oligonucleotides. Very high-resolution separations of oligonucleotides to 60 bases can be obtained. Separations can be performed under non-denaturing or denaturing conditions because the column can be operated routinely at high temperature or high pH. Analytical methods can be directly transferred to larger NucleoPac PA-100 columns for semi-preparative applications. In this report, methods for the analysis of synthetic oligonucleotides under non-denaturing (pH 8.0) and denaturing (pH 12.4) conditions are presented. A gradient separation for machine-grade DNA at pH 8.0 is developed. Transfer of an analytical method to a 22-mm-diameter semi-preparative column is also demonstrated.
一种用于合成寡核苷酸高分辨率分析和纯化的强阴离子交换高效液相色谱柱——NucleoPac PA - 100已被研发出来。可以实现对长达60个碱基的寡核苷酸进行非常高分辨率的分离。由于该色谱柱可在高温或高pH条件下常规运行，因此分离可在非变性或变性条件下进行。分析方法可直接转移至更大的NucleoPac PA - 100色谱柱用于半制备应用。在本报告中，介绍了在非变性（pH 8.0）和变性（pH 12.4）条件下分析合成寡核苷酸的方法。开发了在pH 8.0条件下对工业级DNA的梯度分离方法。还展示了将分析方法转移至直径为22 mm的半制备色谱柱的过程。
