高粱基因组DNA提取方法的比较与优化

Sorghum Genomic DNA Extraction Methods and Optimization of Process

目的:比较CTAB法和高盐低pH法提取高粱总DNA效果,进一步优化建立高盐低pH值法提取高粱基因组DNA的方法。方法:采用CTAB法、高盐低pH法对高粱叶片DNA进行提取,并从电泳结果、纯度、得率三方面进行比较研究。通过提取缓冲液中SDS浓度的优化,不同组织器官的DNA提取纯化以及不同沉淀方法的优化,比较不同方法对高粱基因组DNA提取的影响。结果:高盐低pH法DNA平均得率为689.36μg/g,略小于CTAB法的718.26μg/g,但其A260/A230平均值为1.854,比CTAB法的2.164更接近标准要求。高盐低pH法提取高粱基因组DNA的适宜条件确定如下:提取液中含有2%SDS、2%PVP、2%β-巯基乙醇;水浴时间30min;沉淀方法采用0.7体积异丙醇室温沉淀的沉淀方法。结论:综合考虑高盐低pH法是提取高粱基因组DNA的最佳方法。

Objective：To select the best method to extract DNA from sorghum.Method：As compared the CTAB DNA extraction method with the high-salt low-pH method（HSLPM） from the electrophoresis results,purity and yield.Result：The high-salt low-pH method was better than CTAB method.The extract ratio of CTAB method was higher than high-salt low-pH method while the OD260/280 value of CTAB method was higher than that of the high-salt low-pH method.Conclusion：Compared with CTAB,the high-salt low-pH method was more simple,time saving and with higher extract purity.Considered above all,the high-salt low-pH method was the best method to extract sorghum leaf DNA.