适用于SSR分析的半粒水稻干种子DNA快速提取

Rapid DNA Extraction from Half-grain Rice Dry Seeds for SSR Analysis

为了寻找操作简便、耗时短、成本低,适用于简单重复序列(SSR)分析的水稻基因组DNA快速提取方法,以水稻沈农265的半粒干种子为材料,采用SDS提取液进行一步裂解,并将RNA去除操作融入抽提过程,SDS浓度为0.5%(W/V),水浴10min,氯仿/异戊醇抽提,简单快速地获得了高质量水稻基因组DNA。经琼脂糖凝胶电泳检测,DNA纯度较高,完整性较好,能够满足SSR扩增模板的需求。该方法可大大缩短水稻种子DNA提取时间,为SSR标记在水稻种子遗传多样性分析、分子标记辅助选择等方面的应用奠定基础。

To explore an easy, rapid and economical approach for extracting rice （Oryza sativa L.） genomic DNA for SSR （Simple Sequence Repeat） analysis, half-grain dry seed of rice Shennong265 is taken as tested material, just one step was used for cell lysis by SDS buffer, and the operation of RNA digestion was integrated into protein extraction. During this process, the SDS concn was 0.5% （W/ V）, water bath time was 10min, extraction with cbloroform/isoamyl alcohol was used. High quality genomic DNA from rice seed was obtained simply and quickly. In this paper, the results of agarose gel electrophoresis all showed that the purity and integrity of the isolated DNA were satisfying; the amplified bands were clear as well as good in repeatability and stability, indicating that it can be used in SSR marker analysis. This is a time-saving method for extraction of genomic DNA, and can be effectively applied in genetic diversity analysis of rice seed and marker aided breeding activity.