从甜瓜子叶中提取总DNA

Extraction of total DNA from cotyledons of melon (Cucumis melo L.)

用分子标记方法检测甜瓜杂交种子纯度,如果从真叶中提取DNA,检测周期至少需要15d。为了缩短检测周期,我们分别以甜瓜的成熟干种子、吸胀水后“露白”种子、3d龄刚转绿子叶、6d龄子叶、9d龄子叶和12d龄子叶为材料,进行了提取总DNA的研究。结果表明:除了干种子和吸胀水后“露白”种子外,其他的材料都可以提取到DNA,但是DNA的质量因子叶日龄不同而存在很大的差异。不同日龄的子叶中,以3d龄子叶提取的DNA质量好。子叶6d龄时,提取的DNA质量次于3d龄,少量DNA开始出现降解,以后随着子叶日龄的增加,DNA降解加重。另外,与以真叶为材料提取的DNA样品相比较,子叶DNA样品中的蛋白质等杂质含量高,应增加氯仿抽提纯化次数。

For fast checking of the melon hybrid seeds the purity with molecular markers, the examination at least needs 15 days if the DNA was extracted from true leaf. In this research, for the purpose of shortening examination period, the total DNA was extracted from mature dry seeds, water-soaked-well seeds, different ages of cotyledons and young true leaf of Cucumis melo L., by CTAB method. The result showed that DNA was obtained from all materials except mature dry seeds and water-soaked-well seeds. But DNA quality obtained from different materials was different. The quality of DNA extracted from 3 days old cotyledons was the best. The older of cotyledons, the more serious degradation of DNA extracted. Moreover, compared with DNA sample extracted from young true leaf, the DNA samples from cotyledons had more protein and were impure, the steps of purification with chloroform should be increased.