一种从凝胶中切下的标记限制片段进行滤膜杂交的简单方法。

A simple method for filter hybridization of labeled restriction fragments excised from gels.

作者信息

Marotta R, Bremenkamp M, Salamini F

出版信息

Anal Biochem. 1983 Apr 1;130(1):27-31. doi: 10.1016/0003-2697(83)90645-0.

Abstract

Labeled DNA restriction fragments excised from agarose or bisacrylylcystamine-acrylamide gels can be used for hybridization to nitrocellulose-bound DNA without eliminating the gel matrix. A gel slice containing the labeled fragment is excised, dissolved by heating at 105 degrees C (in the presence of beta-mercaptoethanol for bisacrylylcystamine-acrylamide gels), and added to the hybridization mixture. The presence of agarose or polyacrylamide in the solution does not inhibit hybridization. The method is simple, rapid, and allows complete recovery of the probe.

摘要

从琼脂糖或双丙烯酰胱胺-丙烯酰胺凝胶上切下的标记DNA限制片段可用于与结合在硝酸纤维素上的DNA杂交，而无需去除凝胶基质。切下含有标记片段的凝胶切片，在105℃加热溶解（对于双丙烯酰胱胺-丙烯酰胺凝胶，需在β-巯基乙醇存在下），然后加入杂交混合物中。溶液中琼脂糖或聚丙烯酰胺的存在不会抑制杂交。该方法简单、快速，且能使探针完全回收。