一种快速有效检测SSR标记的非变性聚丙烯酰胺凝胶的银染方法

A rapid and effective method of silver staining for detecting SSR markers in nondenaturing polyacrylamide gels

聚丙烯酰胺凝胶银染法是一种具有高分辨率检测SSR标记的有效方法，然而传统的银染方法存在操作步骤较多、所需时间较长及试剂种类与用量较多等不足．本研究利用菜心和烟草SSR标记为研究对象，建立一种能快速有效检测其PCR扩增产物的非变性聚丙烯酰胺凝胶的银染方法．该方法包括染色和显影2个主要操作步骤，整个银染流程耗时6～7min，只需要AgNO3、NaOH和甲醛3种试剂，所检测SSR标记的DNA条带清晰、易辨．与其他银染法相比，本研究建立的银染法具有易于操作、耗时少、所需试剂种类和用量少及检测效果好的优势．

Silver staining is an effective method to detect SSRs with high resolution in polyacrylamide gel. However the conventional methods of silver staining are time-consuming because they have a number of time-dependent procedures with more kinds of reagents. Here we reported a rapid and effective silver staining method for detecting PCR amplification products in nondenaturing polyacrylamide gel by using SSRs of flowering Chinese cabbage and tobacco. This method includs two major steps-- staining and development, within 6 to 7 rains, and requirs only three reagents which are HNO3 , NaOH and HCOH. With the method, DNA fragments of SSRs are unambiguously detected with high resolution. Compared with other silver staining methods, this new method has advantages of easy operation, time-saving, less amount of chemical reagents and good detection effect.