| 基于PCR的染色体步移中单引物扩增的克服与非特异引物的利用 |
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| 引用本文： | 刘春香,张卫华,孙小镭.基于PCR的染色体步移中单引物扩增的克服与非特异引物的利用[J].中国生物化学与分子生物学报,2010,26(4):380-385. |
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| 作者姓名： | 刘春香 张卫华 孙小镭 |
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| 作者单位： | 1.潍坊学院（现在：山东省农科院在职博士后）;2.山东省农科院蔬菜所; |
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| 基金项目： | 山东省博士后择优资助项目(No.200603041,No.30471975); 山东省优秀科学家奖励基金(No.2007BSC02701)资助~~ |
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| 摘 要： | 基于PCR的染色体步移(PCR-Walking)方法已有许多种,包括反向PCR、连接介导的PCR、随机引物PCR等.在众多的方法中,经常存在由通用引物引起的单引物非特异扩增现象.本文综述了连接介导的PCR-Walking中单引物扩增的形成原理及克服方法.克服单引物扩增主要是使接头引物在DNA两端的接头上只有1个结合位点,从而避开单引物扩增.常用的方法有3′端加氨基修饰的不对称接头、泡泡状接头或Y字型接头及单寡核苷酸接头等方法.还介绍了2种利用通用引物非特异扩增克隆目的序列的方法:引物错配法及基于RAPD原理的单引物PCR法.
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| 关 键 词： | 单引物扩增 PCR 染色体步移 |
| 收稿时间： | 2009-9-23 |
Applying Random Primer and Overcoming Single Primer Amplification in PCR-based Chromosome Walking |
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| LIU Chun-Xiang,ZHANG Wei-Hua,SUN Xiao-Lei.Applying Random Primer and Overcoming Single Primer Amplification in PCR-based Chromosome Walking[J].Chinese Journal of Biochemistry and Molecular Biology,2010,26(4):380-385. |
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| Authors: | LIU Chun-Xiang ZHANG Wei-Hua SUN Xiao-Lei |
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| Institution: | ( Institute of Vegetables, Shandong Academy of Agricultural Science, Jinan 250100,China;
Bio-Tech Engineering College of Weifang University, Weifang 261061, Shandong, China) |
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| Abstract: | Abstract: There are many chromosome walking methods based on PCR, include inverse PCR, ligation based PCR walking, random primer based PCR walking. Unspecific amplification caused by general single primer always lies in these methods. In this paper, the theory of single primer amplification formed, overcoming methods of single primer amplification, and the usage of unspecific amplification by general primer were summarized. The major introducing part is the overcoming methods of single primer amplification. |
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| Keywords: | single primer amplification PCR chromosome walking |
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