Budelier K, Schorr J
QIAGEN, Inc., Valencia, California, USA.
Curr Protoc Mol Biol. 2001 May;Chapter 2:Unit2.1B. doi: 10.1002/0471142727.mb0201bs42.
Column chromatography has evolved to provide a rapid and effective alternative to more laborious methods for preparing high-quality DNA, such as CsCl-gradient centrifugation. This unit describes the use of a column made of a unique anion-exchange resin that selectively binds nucleic acids, allowing rapid separation of DNA from contaminating RNA, proteins, carbohydrates, and metabolites. The procedure employs columns supplied by QIAGEN; other preparation methods are available from other suppliers. A crude nucleic acid sample (usually a cleared cell lysate) is applied to the QIAGEN tip under conditions that favor binding. Contaminants in the sample are washed from the column with a moderate-salt buffer, and DNA is eluted using a high-salt buffer.
柱色谱法已发展成为一种快速有效的方法，可替代诸如氯化铯梯度离心等更为繁琐的制备高质量DNA的方法。本单元介绍了一种由独特的阴离子交换树脂制成的柱子的使用方法，该树脂能选择性地结合核酸，从而使DNA能快速从污染的RNA、蛋白质、碳水化合物和代谢产物中分离出来。该程序使用的是QIAGEN公司提供的柱子；其他供应商也有其他制备方法。将粗核酸样品（通常是澄清的细胞裂解物）在有利于结合的条件下加到QIAGEN柱头上。用中等盐浓度的缓冲液从柱上洗去样品中的污染物，然后用高盐缓冲液洗脱DNA。
