小麦基因组DNA快速提取及SSR标记PAGE的银染检测

为了提高SSR-PCR速率,降低试验成本,以8份抗条锈小麦幼苗为材料,比较分析了两种DNA提取方法(CTAB法和SDS法)以及两种SSR标记PAGE银染法(Sanguinetti银染法和Bassam银染法).结果表明:用CTAB法和SDS法均可获得质量较好的DNA,但CTAB法花费的时间较长,提取效率低(在相同条件下SDS法提取速度约为CTAB法的2倍);Sanguinetti银染法染色背景较浅、条带清晰、分辨率高、具有较高的多态性检出率,并且在速度上优于Bassam法,仅需30多min,而Bassam银染法染色步骤烦琐,对试剂质量和水质要求均较高,且未能染色成功.

DNA extraction and SSR PAGE electrophoresis silver staining for wheat seedlings

In order to screen an effective method of DNA extraction and SSR marker silver staining for 8 wheat seedlings,two methods of extraction CTAB and improved-SDS,silver staining Sanguinetti and Bassam were compared.The results indicated that both DNA extraction methods of CTAB and improvedSDS could produce good quality DNA,but CTAB took as long as twice time than improved-SDS.For silver staining Sanguinetti,its background color was light,strap clear and resolution high,and it had higher ratio of polymorphism and took only 30 seconds,much faster than Bassam which procedure was trivial and required quality reagent and water.