| 陈山红心杉基因组DNA提取方法的比较与分析 |
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| 引用本文： | 伍艳芳,徐海宁,肖复明,熊振宇,江香梅.陈山红心杉基因组DNA提取方法的比较与分析[J].江西农业大学学报,2012,34(3):517-521,527. |
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| 作者姓名： | 伍艳芳 徐海宁 肖复明 熊振宇 江香梅 |
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| 作者单位： | 江西省林业科学院,江西省植物生物技术重点实验室,江西南昌330032 |
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| 基金项目： | 国家林业公益性行业科研专项 |
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| 摘 要： | 采用尿素法、柠檬酸钠法、改良的CTAB法和SDS法4种方法提取陈山红心杉基因组DNA,并对提取效果进行比较和分析。结果表明,改良的CTAB法较适合陈山红心杉基因组DNA提取,其提取的DNA纯度高、质量好,具有典型的天然DNA分子的标准紫外吸收光谱特点,A260/A280比值在1.8左右,且凝胶检测条带整齐清晰,杂质少、无明显降解。而尿素法和柠檬酸钠法难以提取基因组DNA,SDS法因提取的DNA有蛋白质、多糖等杂质污染均不太适用陈山红心杉基因组DNA提取。酶切分析也证明,改良的CTAB法提取的DNA适用于进行后续分子生物学分析。采用改良的CTAB法能够提取得到陈山红心杉幼叶、老叶、嫩芽和幼茎的基因组DNA产率不同,分别为158.4μg/g、129.6μg/g、177.6μg/g和98.40μg/g。考虑到幼叶样品的采集要比嫩芽方便,因此,可以直接采用幼叶进行陈山红心杉基因组DNA提取。
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| 关 键 词： | 陈山红心杉 DNA提取方法 改良的CTAB法 SDS法 |
Comparative Research on Isolation and Purification Method for Genomic DNA of Cunninghamia lanceolata |
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| WU Yan-fang
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XU Hai-ning
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XIAO Fu-ming
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XIONG Zhen-yu
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JIANG Xiang-mei.Comparative Research on Isolation and Purification Method for Genomic DNA of Cunninghamia lanceolata[J].Acta Agriculturae Universitis Jiangxiensis,2012,34(3):517-521,527. |
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| Authors: | WU Yan-fang XU Hai-ning XIAO Fu-ming XIONG Zhen-yu JIANG Xiang-mei |
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| Institution: | (Jiangxi Provincial Bio-tech Key Lab.for Plant,Jiangxi Academy of Forestry Sciences,Nanchang 330032, China) |
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| Abstract: | The DNA samples of Cunninghamia lanceolata were prepared by Carbamide method,Sodium citrate method,the modified CTAB method and SDS method,respectively.The results showed that improved CTAB method was suitable for extracting the genomic DNA from C.lanceolata.DNA samples obtained by this method possessed the standard ultraviolet absorbance spectrum of pure natural DNA,whose A260/A280 was 1.80 or so,degradation was light and impurity was little.However,Carbamide method and Sodium citrate method could not extract the genomic DNA and SDS method was difficult to get rid of polyhexose completely. Therefore,the three methods were not suitable for extracting the genomic DNA.When tested through the restriction enzyme analysis,the DNA samples prepared by the modified CTAB method could meet the next experiment requirement perfectly.The yield of DNA of young leaves,old leaves,young shoots and young stems obtained by the improved CTAB method were 158.4μg/g,129.6μg/g,177.6μg/g and 98.40μg/g.Young leaves could directly use to extract the genomic DNA after taking the convenience of sample collecting into account. |
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| Keywords: | Cunninghamia lanceolata genomic DNA extraction improved CTAB method SDS method |
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