三种赤潮微藻基因组DNA快速制备方法的研究

采用超声波法、煮沸法和微波法3种方法分别对塔玛亚历山大藻、环状异帽藻和角毛藻进行细胞破碎及快速制备基因组DNA的研究。通过细胞计数和DNA浓度测定的手段对三种方法进行了比较,以选择适合不同藻种的细胞破碎方法。结果表明,塔玛亚历山大藻和环状异帽藻用超声波法破碎效果较好;角毛藻用微波法较好。对用该三种方法制备的基因组DNA做了PCR扩增,电泳检测表明,与CTAB法扩增效果一样。本文建立的微藻DNA快速制备方法有望应用在赤潮藻类的快速分子鉴定方面。

Reliable and rapid genomic DNA extraction method of three species of red tide algae

Recovering DNA of high quality and quantity is a prerequisite for ensuring suitable applications such as detecting red tide algae based on polymerase chain reaction (PCR). We report here DNA rapid isolation protocols for three species of red tide alage: Alexandrium tamarense , Heterocapsa circularisquama and Chaetoceros sp.. The cell walls of algae were crushed by ultrasonication, boiling and microwave method, respectively. And crushing efficiency was calculated by counting cells and assaying yield ratio of genomic DNA. These assays indicated that ultrasonication was an appropriate method for disrupting cell walls of A. tamarense and H. circularisquama , and microwave for Chaetoceros sp.. PCR amplification was made on the extracted DNA got by three methods, and the result showed the same as that by CTAB method. Our DNA extraction protocols can be applied with success to other red tide microalgae and suitable for further molecular analysis.