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坛紫菜(Porphyra haitanensis)叶绿体DNA的快速提取

A Method of Rapidly Extracting Chloroplast DNA from Porphyra haitanensis

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【作者】 邓元告左正宏陈奕欣郑微云

【Author】 DENG Yuangao1,ZUO Zhenghong1, CHEN Yixin1*,ZHENG Weiyun2(1.School of Life Sciences,2.Environmental Science ResearchCenter,Xiamen University,Xiamen 361005,China)

【机构】 厦门大学生命科学学院厦门大学环境科学研究中心 福建厦门361005福建厦门361005福建厦门361005

【摘要】 以阴干的坛紫菜叶状体为材料,通过酶解等方法获取完整的叶绿体,再裂解叶绿体并酚仿抽提获得叶绿体DNA.限制性内切酶HindⅢ和EcoRⅠ双酶切及RAPD检测结果显示:叶绿体DNA与基因组DNA的酶切图谱及RAPD图谱均有比较明显的差异.多次重复实验证明,按此法提取的叶绿体DNA,其得率稳定,并可用作PCR扩增的模板及酶切图谱的构建等.该方法操作简便,过程快捷,可作为大型海藻叶绿体DNA提取的参考方法.

【Abstract】 A convenient method for rapid isolation of chloroplast DNA from Porphyra haitanensis was presented. First, the chloroplasts were isolated, and then lysed in sarcosylproteinase K solution, after that the chloroplast DNA was extracted and purified by phenolchloroform method. The result showed that the chloroplast DNA had different bands from the genomic DNA when double digested with restriction enzyme EcoRⅠ and HindⅢ or RAPD tested. The quality and quantity of the obtained chloroplast DNA was steady and credible by several repeated experiments. It could be used as the template of PCR and construction of physical map. Even more, this method could be the reference method when preparing other algal chloroplast DNA.

【关键词】 坛紫菜叶绿体DNA基因组DNA
【Key words】 Porphyra haitanensischloroplast DNAgenomic DNA
【基金】 科技部海洋863项目(2002A603023);福建省重大农业科技项目(2001Z017)资助
  • 【文献出处】 厦门大学学报(自然科学版) ,Journal of Xiamen University(Natural Science) , 编辑部邮箱 ,2003年06期
  • 【分类号】Q943
  • 【被引频次】9
  • 【下载频次】392
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