Lawson T G, Regnier F E, Weith H L
Anal Biochem. 1983 Aug;133(1):85-93. doi: 10.1016/0003-2697(83)90225-7.
A rapid and effective method for separating oligonucleotides using high-performance ion-exchange chromatography is described. Columns were prepared by adsorbing a layer of polyethylene imine on 5-micron-diameter porous silica followed by crosslinking with a multifunctional oxirane. This weak anion-exchange matrix was found to be useful in the separation of mononucleotides and oligonucleosides containing up to 20 residues in either homo- or heteropolymers. Small analytical columns (4.2 X 150 mm) had a capacity ranging from less than a microgram in analytical separations to several milligrams in the preparative mode. The columns have proven to be especially useful in assessing the purity of precursor blocks, monitoring the chemical synthesis of oligonucleotides, and isolating reaction products after the synthesis.
本文描述了一种使用高效离子交换色谱法分离寡核苷酸的快速有效方法。通过在直径为5微米的多孔硅胶上吸附一层聚乙烯亚胺，然后与多功能环氧乙烷交联来制备色谱柱。发现这种弱阴离子交换基质可用于分离同聚物或杂聚物中含多达20个残基的单核苷酸和寡核苷。小型分析柱（4.2×150毫米）的容量在分析分离中小于1微克，在制备模式下可达几毫克。这些色谱柱已被证明在评估前体模块的纯度、监测寡核苷酸的化学合成以及合成后分离反应产物方面特别有用。
