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本研究以柱花草(Stylosanthes guianensis)不育和可育两种分离群体为试验材料,采用优化改进后的CTAB法提取柱花草不育和可育两种分离群体的叶片的基因组DNA。试验结果表明:提取出的DNA检测浓度值373.7~604.3 ng/μL,A260/A280处于1.83~1.91,A260/A230处于1.90~2.05。所提的DNA琼脂糖凝胶的电泳条带显示清晰明亮,没有明显的拖带现象。ISSR-PCR琼脂糖凝胶电泳在不育和可育植株的条带之间具有明显的多态性条带。说明用此优化改进的CTAB法提取的目标基因组DNA的纯度高,质量好。能很好地用于分子遗传特性分析和以ISSR-PCR和SRAP-PCR为基础的分子生物学方面。该改进的试验方法简单、高效、快捷。
In this study, Stylosanthes guianensis male sterile and fertile two populations were used as experimental materials, and genomic DNA was extracted from the leaves of Sterile and Fertile populations of Stylosanthes guianensis using optimized and improved CTAB method. The results showed that the detected concentration of DNA was 373.7 ~ 604.3 ng / μL, A260 / A280 was 1.83-1.91, A260 / A230 was 1.90-2.05. The DNA bands of the DNA agarose gel displayed clear and bright, there is no obvious dragging phenomenon. ISSR-PCR agarose gel electrophoresis showed significant polymorphic bands between the bands of sterile and fertile plants. It shows that the target genomic DNA extracted by this optimized and improved CTAB method has high purity and good quality. It is well suited for molecular genetic characterization and molecular biology based on ISSR-PCR and SRAP-PCR. The improved test method is simple, efficient and fast.