| 一种快速·有效分离T-DNA插入位点的侧翼序列的方法——DW-ACP-PCR技术 |
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| 引用本文： | 王葵娣,郑服丛.一种快速·有效分离T-DNA插入位点的侧翼序列的方法——DW-ACP-PCR技术[J].安徽农业科学,2007,35(28):8817-8818,8841. |
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| 作者姓名： | 王葵娣 郑服丛 |
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| 作者单位： | 中国热带农业科学院环境与植物保护研究所,海南儋州,571737;中国热带农业科学院环境与植物保护研究所,海南儋州,571737 |
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| 基金项目： | 国家自然科学基金（30260046）. |
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| 摘 要： | DW-ACP-PCR(DNAWalking-Annealing control primer-PCR)是一种分离已知DNA序列的侧翼未知序列的新技术。该技术通过3个嵌套的特异引物分别和ACPC复性控制引物组合,进行3步连续的PCR反应,所有反应可以在1 d内完成。ACP的特殊结构能有效地控制非特异扩增。用DW-ACP-PCR技术扩增7个稻瘟菌T-DNA插入突变体的T-DNA插入位点的侧翼序列,均获得了特异目标片段。
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| 关 键 词： | DW-ACP-PCR T-DNA定位 侧翼序列 稻瘟菌 |
| 文章编号： | 0517-6611（2007）28-08817-02 |
| 修稿时间： | 2007-05-09 |
A Rapid and Efficient Method of Isolating the Flanking Sequence of T-DNA Inserted Site-DW-ACP-PCR Technique |
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| WANG Kui-di et al.A Rapid and Efficient Method of Isolating the Flanking Sequence of T-DNA Inserted Site-DW-ACP-PCR Technique[J].Journal of Anhui Agricultural Sciences,2007,35(28):8817-8818,8841. |
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| Authors: | WANG Kui-di |
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| Institution: | Institute of Environment and Plant Protection;CATAS;Danzhou;Hainan 571737 |
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| Abstract: | DW-ACP-PCR(DNA Walking Annealing control primer PCR) was a new technique of isolating the unknown flanking sequences in known DNA sequence,in which 3 nested specific primers were resp.combined with ACPC renaturation control primers for 3 step continuous PCR reactions.All the reactions needed only one day to be accomplished.The unique structure of ACP could control nonspecific amplification effectively.It was introduced that DW-ACP-PCR technique was used to amplify the flanking sequences of T-DNA inserted site of 7 T-DNA inserted mutants of Magnaporthe grisea. |
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| Keywords: | DW-ACP-PCR T-DNA location Flanking sequences Magnaporthe grisea |
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