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Basic Methods in Cellular and Molecular Biology
Purifying DNA From Agarose Gels
Description
Purifying DNA from agarose gels lets students recover a separated DNA fragment after electrophoresis. The DNA is first isolated from the gel, then moved through a silica column for cleanup.
The process has three main steps: binding, washing, and eluting. DNA is believed to bind to silica when hig...
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Transcript
胶纯化是用来在琼脂糖凝胶中回收电泳分离的DNA片段的标准步骤。将胶溶解后，让它通过一种特殊的滤膜，这一步骤使得DNA与其他杂质，如盐，游离核苷酸，酶等分离开，从而可以进行下游的应用操作。
从琼脂糖凝胶中回收DNA的基本原理涉及到一系列的结合，清洗和洗脱步骤。当胶完全溶解在溶解缓冲液中后，上样到一个"离心柱"上，它通过离心可使DNA分子特异性的结合到硅胶膜上，而其他的杂质则通过柱子到了收集管。
由于凝胶溶解缓冲液中的高盐浓度，使得DNA可以结合到硅胶上。该缓冲液会破坏膜周围的水合结构，在强阴性的膜和阴性的DNA之间建立一个阳离子盐桥。剩余的杂质则被乙醇清洗去掉。
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