| 一种DNA提取新方法在转基因水稻种子检测中的应用 |
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| 引用本文： | 张富丽,刘勇,宋君,雷绍荣,郭灵安,王东,尹全,刘文娟,常丽娟.一种DNA提取新方法在转基因水稻种子检测中的应用[J].湖北农业科学,2012,51(10):2118-2120. |
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| 作者姓名： | 张富丽 刘勇 宋君 雷绍荣 郭灵安 王东 尹全 刘文娟 常丽娟 |
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| 作者单位： | 1. 四川省农业科学院,分析测试中心,成都610066
2. 四川省农业科学院,分析测试中心,成都610066;四川省农业科学院,植物保护研究所,成都610066 |
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| 基金项目： | 农业部转基因重大专项(2011ZX08011-006);四川省农业科学院青年基金项目(2010QNJJ-031) |
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| 摘 要： | 将水稻种子打磨成粉,取100 mg粉末加入SDS和NaC1等试剂配制成高盐提取液,然后提取基因组DNA.超微量分光光度计NanoDrop 1000检测及Eco RV酶切反应的结果表明,该方法比CTAB法的DNA浓度更高、纯度更好.以提取得到的DNA为模板,采用PCR法分别对水稻特异性内源基因SPS及转基因水稻外源基因Bt进行扩增,均能扩增到清晰的目标条带.这种DNA提取新方法不仅简单快速、成本低廉,而且提取到的DNA质量高,扩增效果好,可有效地用于水稻种子转基因成分的检测.
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| 关 键 词： | DNA提取 高盐 转基因水稻 种子 |
A New DNA Extracting Approach for Detection of Transgenic Rice Seeds |
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| ZHANG Fu-li
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LIU Yong
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SONG Jun
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LEI Shao-rong
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GUO Ling-an
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WANG Dong
,
YIN Quan
,
LIU Wen-juan
,
CHANG Li-juan.A New DNA Extracting Approach for Detection of Transgenic Rice Seeds[J].Hubei Agricultural Sciences,2012,51(10):2118-2120. |
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| Authors: | ZHANG Fu-li LIU Yong SONG Jun LEI Shao-rong GUO Ling-an WANG Dong YIN Quan LIU Wen-juan CHANG Li-juan |
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| Institution: | a(a.Analysis and Determination Center,b.Plant Protection Research Institute,Sichuan Academy of Agricultural Sciences, Chengdu 610066,China) |
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| Abstract: | 100 mg rice seeds powder was added into high salt solution composed of SDS and NaCl to extract the genomic DNA.According to ultra-micro spectrophotometer NanoDrop 1000 test and Eco RV digestion,this DNA extracting method was better than CTAB due to its higher yield and better quality of DNA.The endogenous gene SPS and foreign Bt gene could be amplified with distinct and specific target bands by PCR using the obtained DNA as template.This method was fast,simple,economy and resultful,thus could be used for ingredient detection of transgenic rice seeds. |
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| Keywords: | DNA extraction high salt transgenic rice seed |
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