枸杞花药线粒体分离纯化方法

A Method of Extracting and Purifying Anther Mitochondria in Lycium barbarum L.

为了获得高质量的枸杞花药线粒体,本文介绍了一种差速离心分离纯化枸杞花药线粒体的方法。通过Jannus Green B显色反应、线粒体跳动特性以及电镜观察鉴定,结果表明采用本差速离心法提取的线粒体纯度高、活性强、结构完整。进一步通过琼脂糖电泳、光密度值及核特异引物PCR检测表明,由该线粒体裂解的mtDNA不仅纯度高、质量好,而且得率高。本差速离心法具有简捷、高效和经济等优点。用该方法获得的线粒体完全能够满足线粒体基因组、蛋白质组等相关分子生物学研究要求。

In order to obtain high quality anther mitochondria, this paper introduces a method of extracting and purifying anther mitochondria in Lycium barbarum L. by differential centrifugation. The results showed that mito- chondria of using the method extracting are pure, strong activity and complete structure by the Jannus Green B staining, beating characteristics of mitochondria and identification of electron microscopy. And furthermore results showed that mtDNA form the mitochondria not only are high purity and good quality, but also high yield through the detecting of the agarose gel electrophoresis, optical density and nuclear specific primer PCR. It indicates that this method has easy, quick, efficient and economic advantages. And the mitochondria of using the method extracting can completely meet the research requirements of the mitochondrial genome, proteome and other related molecular biology experiment.