A Method of Rapidly Extracting Chloroplast DNA from Porphyra haitanensis
- 环境生态－已发表论文 
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.A convenient method for rapid isolation of chloroplast DNA from Porphyra haitanensis was presented. First, the chloroplasts were isolated, and then lysed in sarcosylproteinase K solution, after that the chloroplast DNA was extracted and purified by phenolchloroform method. The result showed that the chloroplast DNA had different bands from the genomic DNA when double digested with restriction enzyme EcoRⅠ and HindⅢ 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.