| 适合于基因组测序的红麻高纯度线粒体DNA提取 |
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| 引用本文： | 赵杨,陈鹏,周瑞阳. 适合于基因组测序的红麻高纯度线粒体DNA提取[J]. 东北农业大学学报, 2011, 42(4): 98-101 |
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| 作者姓名： | 赵杨 陈鹏 周瑞阳 |
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| 作者单位： | 广西大学农学院,南宁,530004;广西大学农学院,南宁,530004;广西大学农学院,南宁,530004 |
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| 基金项目： | 国家自然科学基金项目,教育部科学技术研究重点项目 |
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| 摘 要： | 采用红麻黄化苗为试材,结合密度梯度离心和差速离心的方法提取线粒体DNA(mtDNA)以满足测序要求.结果表明,蔗糖密度梯度离心法比Percoll密度梯度离心法更适合于红麻线粒体的分离.分离后的线粒体经DNaseI消化核DNA,并采用Jannus绿检测线粒体的完整性,SDS和蛋白酶K裂解线粒体,并用酚/氯仿抽提除去蛋白,...
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| 关 键 词： | 红麻 线粒体 mtDNA 测序 |
Extraction of high purity mitochondrial DNA from kenaf for genome sequencing |
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| ZHAO Yang,CHEN Peng,ZHOU Ruiyang. Extraction of high purity mitochondrial DNA from kenaf for genome sequencing[J]. Journal of Northeast Agricultural University, 2011, 42(4): 98-101 |
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| Authors: | ZHAO Yang CHEN Peng ZHOU Ruiyang |
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| Affiliation: | (College of Agriculture,Guangxi University,Nanning 530004,China) |
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| Abstract: | An efficient method for extraction of mitochondrial DNA(mtDNA) from etiolated tissues of kenaf for sequencing was developed.The results showed that sucrose gradient centrifugation was better than Percoll’ s as for mitochondria extraction of kenaf.The mitochondria was digested with DNase I to remove the nuclear DNA,then was extracted with phenol/chloroform,followed by ethanol deposition,and got rid of RNA contamination by treating RNase A.The mtDNA was detected by gelose electro-phoresis and ultraviolet spectrophotometer,furthermore,a chloroplast and nuclear-specific primers were designed to detect mtDNA purity.The results showed that the mtDNA extracted in this way had high purity and no contamination,then could be used as genome sequencing. |
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| Keywords: | mtDNA |
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