A rapid and effective method for silver staining of PCR products separated in polyacrylamide gels
A rapid and effective method for silver staining of PCR products separated in polyacrylamide gels
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DOI：
10.1002/elps.201400182
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发表时间：
2014-09-01
期刊：
影响因子：
2.9
通讯作者：
Shi, Shengyou
中科院分区：
文献类型：
--
作者：
Liang, Qingzhi;Wen, Dingqing;Shi, Shengyou
随着分子数量遗传学，特别是遗传连锁图谱构建、数量性状基因座定位或基因精细定位和关联分析等的发展，越来越多的聚丙烯酰胺凝胶中分离的PCR产物需要进行银染。然而，传统的银染色程序复杂且耗时，因为它们需要在使用前对多种溶液进行大量准备和处理。在本研究中，开发了一种简单快速的 PCR 产物银染方案。与常规方案相比，减少了步骤数，从而在7分钟内实现了PCR产物的检测，节省了时间和资源。目前染色流程中的固定染色液和显色液分别可重复利用12次和8次，大大降低了成本。同时，改进后的方法灵敏度显着提高，在变性聚丙烯酰胺凝胶中最低可检出0.097 ng/L的DNA量。本研究开发的方案将促进分子定量遗传学的发展。
With the development of molecular quantitative genetics, particularly, genetic linkage map construction, quantitative trait loci mapping or genes fine mapping and association analysis etc., more and more PCR products separated in polyacrylamide gels need to be silver-stained. However, conventional silver-staining procedures are complicated and time-consuming as they require a lot of preparation and handling of several solutions prior to use. In this study, a simple and rapid protocol for silver staining of PCR products was developed. The number of steps was reduced compared to conventional protocols, thus achieving detection of PCR products in 7 min, saving time and resources. Fixation and staining solution and developing solution in present staining procedure allowed a reutilization for 12 and 8 times, respectively, reducing the cost greatly. Meanwhile, the sensitivity was significantly improved with the improved method and the minimum of 0.097 ng/L of DNA amount can be detected in denaturing polyacrylamide gel. The protocol developed in this study will facilitate the development of molecular quantitative genetics.