| 小麦中转基因成分PCR与实时荧光PCR的定性检测低限 |
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| 引用本文： | 白月,栾凤侠,王珣.小麦中转基因成分PCR与实时荧光PCR的定性检测低限[J].麦类作物学报,2009,29(2). |
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| 作者姓名： | 白月 栾凤侠 王珣 |
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| 作者单位： | 1. 黑龙江出入境检验检疫局,黑龙江哈尔滨,150001
2. 黑龙江省农业科学院生物技市研究所,黑龙江哈尔滨,150086 |
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| 基金项目： | 国家质检总局科研项目，科技部社会公益研究专项重点项目子课题，国家标准委下达的国家标准制定项目 |
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| 摘 要： | 为了确定小麦转基因成分PCR和实时荧光PCR方法的定性检测低限,将转基因小麦B73与非转基因小麦(检测外源基因)、小麦与大米(检测内源基因)分剐进行质量分数配比后提取DNA用于测定相对检测低限,再将100%转基因小麦B73的DNA进行浓度稀释用于测定绝对检测低限,并应用已知的NOS、bar、ubiquitin,ui-dA(GUS)外源基因和肌Wx012、GAG56D内源基因的引物和探针时模板DNA分别进行PCR与实时荧光PCR扩增.结果表明,最终确定PCR方法检测小麦转基因成分的相对检测低限为0.1%(质量分数),绝对检测低限为0.5 ng/ⅡL;实时荧光PCR方法的相对检测低限为0.1%(质量分数),绝对检测低限为0.01 ng/μL.所确定的检测低限可满足国家对转基因产品的最低标识要求.
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| 关 键 词： | 转基因小麦 实时荧光PCR 绝对检测低限 相对检测低限 |
Detection Limits of PCR and Realtime PCR for Genetically Modified Components in Transgenic Wheat |
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| BAI Yue,LUAN Feng-xi,WANG Xun.Detection Limits of PCR and Realtime PCR for Genetically Modified Components in Transgenic Wheat[J].Journal of Triticeae Crops,2009,29(2). |
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| Authors: | BAI Yue LUAN Feng-xi WANG Xun |
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| Institution: | 1.Heilongjiang Entry-Exit Inspection and Quarantine Bureau;Harbin Heilongjiang 150001;China;2.Biotechnology Research Center;Heilongjiang Academy of Agricultural Sciences;Harbin;Heilongjiang 50086;China |
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| Abstract: | This study is on the detection limits of PCR and realtime PCR for genetically modified components in wheat.B73 was the transgenic wheat line which contained ubiquitin promotor,NOS terminate,bar and uidA(GUS) reported gene,Wx012 and GAG56D endogenesis gene.Genomic DNA from transgenic wheat and non tansgenic wheat,wheat and rice(wt/wt) were extracted for the relative detection.B73 genamic DNA was tenfold serially diluted to define the absolute detection limit.After amplifing the endogenesis gene and extrinsic... |
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| Keywords: | PCR |
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