新生隐球菌线粒体的快速抽提和电镜观察

Rapid Extraction of MtDNA from Cryptococcus neoformans and Examined by Electronic Microscopy

通过NovoZym2 34酶溶壁和低渗机械振荡破壁相结合 ,应用差速离心法分离并纯化了对数生长期的新生隐球菌线粒体 ,然后从经DNaseI处理的线粒体制备液中分离纯化线粒体DNA ;并对差速离心中所获得的菌体、原生质体、线粒体三部分沉淀进行了透射电镜观察 ,结果均证明了我们所抽提的DNA是纯净的 ,适用于酶切分析和PCR分析研究 ,由此成功地建立了快速有效分离和纯化线粒体DNA的方法。

Cryptococcus neoformans may be grown to the exponential phase,are broken by a combination of NovoZym234 and mechanical means,and mitochondrial DNA was extracted from DNaseI-treated mitrochondrial preparetion by differential centrifugation.Three pellets,including yeast cell,protoplasts,mitochondrial,were examined by transmission electronic microscopy.The resulting mtDNA is sufficicently pure for restriction endonucleases analysis and PCR in further studying.A rapid and effective method for the preparation of the mtDNA of C.neoformans was established.