利用SSR分子标记快速鉴定杂交水稻种子纯度技术体系的优化

针对目前SSR分子标记技术快速鉴定杂交水稻种子纯度方法商业化应用的限制因素,从DNA提取、PCR反应体系和程序及凝胶电泳等方面进行了技术体系的优化。原方法经优化后,DNA提取时间从每次5h以上减少到2.5min,PCR反应过程从200min缩短至65min,改聚丙烯酰胺凝胶电泳为琼脂糖凝胶电泳,电泳时间由3.0-3.5h缩短为1h,且通过减少dNTP、Taq酶的用量,对琼脂糖凝胶反复利用,大大降低了成本,减少了废弃物的污染,从而建立了准确、简单、快速、成本低廉的快速鉴定杂交稻种子纯度的技术体系。

Optimization of the Rapid Purity Identification System of Hybrid Rice Seeds by Using SSR Markers

To overcome the limiting factors in the commercialization of the existing rapid purity identification system of hybrid rice seeds by using SSR markers, the studies were conducted on the improvement of the technology system in the aspects of DNA extraction, PCR amplification procedure and gel electrophoresis. After optimization, the time for DNA extraction, PCR amplification and gel electrophoresis was decreased from 300, 200 and 180 -210 minutes to 2.5, 65 and 60 minutes, respectively. Meanwhile, the cost was greatly reduced and the pollution to the environment was abated by cutting down the dosage of dNTP and Taq DNA polymerase, and reusing the gelose gel. Therefore, an optimized accurate, simple, rapid and low costing technical system of fast purity identification of hybrid rice seeds was established.