甘蓝型油菜成熟籽粒DNA快速提取方法探讨

Study of Rapid Methods for Isolation of DNA from the Seeds of Brassica napus L.

以室温保存不同年份的甘蓝型油菜种子为材料,采用改良的CTAB法和DNA抽提试剂盒分别提取甘蓝型油菜籽粒DNA,探讨适合甘蓝型油菜籽粒DNA提取的方法。结果表明,不同保存年份种子提取的DNA含量无显著差异;受种皮色泽的影响也较小;用改良的CTAB法比试剂盒所提DNA产量高,结构完整性好,成本较低。PCR扩增检测结果表明两种方法获得的DNA均能满足一般的分子实验。因此,从油菜干籽粒中直接提取DNA可以节约时间,显著提高工作效率,为快速高效地进行甘蓝型油菜种质资源和遗传多样性分析奠定了基础,同时,也利于快速鉴定推广品种是否带有外源基因。

Based on the methods of modified CTAB and EasyPure Plant Genomeic DNA Extraction Kit, genomic DNA was extracted from the dry seeds of different storage years to evaluate the DNA extraction methods from seed of Brassica napus L. The results showed that the concentration of extracted DNA had no significant difference across the storage time of this set of seeds. The genomic DNA contents had also no significant difference between the yellow seed and black seed. Compared with Extraction Kit, the genomic DNA isolated by modified CTAB method had higher concentration, better integrity and lower cost. The quality of DNA was proved perfect for ordinary molecular experiments through polymerase chain reaction. DNA extraction from seed significantly improved the ＇work efficiency. This is useful in germplasm identification and genetic diversity studies, as well as rapidly identification of cuhivars with foreign genes.