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一种改进的丝状真菌DNA提取方法

A Modified Method for Isolating DNA from Fungus

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【作者】 张颖慧魏东盛邢来君李明春

【Author】 ZHANG Ying-Hui WEI Dong-Sheng XING Lai-Jun LI Ming-Chun* (Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin 300071)

【机构】 南开大学微生物学系分子微生物学与技术教育部重点实验室南开大学微生物学系分子微生物学与技术教育部重点实验室 天津300071天津300071

【摘要】 以丝状真菌雅致枝霉(Thamnidium elegans)和深黄伞形霉(Umbelopsis isabellina)为材料,使用优化的CTAB法提取基因组DNA。改进后的方法使用液氮冻融以及玻璃珠振荡的方法代替了传统的液氮研磨,所需菌体量少,而得到的基因组DNA比用传统的CTAB法得到的基因组DNA产率高、纯度好、且步骤简单,适用于一次微量提取多个样品的基因组DNA。这种方法得到的基因组DNA可用于大部分分子生物学基本实验如PCR和DNA的酶切等。

【Abstract】 Genomic DNA of two fungi Thamnidium elegans and Umbelopsis isabellina were extracted with an amended Cetyltrimethyl Ammonium Bromide (CTAB) method. This modified method uses repeated freezing in liquid nitrogen and thawing with combination of shocking with glass beads to replace of the tra- ditional method. Quality and concentration of DNA extracted by the modified methodwere tested. Compared with the traditional method, higher yield and purity of genomic DNA were obtained with less amount ofmy- celium. The result indicted that this is a simple and highly efficient method, which is suitable to treat many samples at one time and for basic molecular experiments, such as restriction endonuclease reaction and PCR.

【基金】 天津市科技支撑计划重点项目(已批准,编号待定)
  • 【文献出处】 微生物学通报 ,Microbiology , 编辑部邮箱 ,2008年03期
  • 【分类号】Q93
  • 【被引频次】127
  • 【下载频次】2037
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