| 一种用PUC质粒高效克隆PCR产物的方法 |
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| 引用本文： | 叶军,孔杰.一种用PUC质粒高效克隆PCR产物的方法[J].海洋水产研究,1995,16(1):72-75. |
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| 作者姓名： | 叶军 孔杰 |
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| 作者单位： | [1]青岛海洋大学海洋生命学院 [2]中国水产科学研究院黄海水产研究所 |
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| 摘 要： | ＰＵＣ质粒用Ｓａｍｌ酶切，经Ｔａｑ酶和ｄＴＴＰ处理，构建成ｄＴ载体。ＰＣＲ产物经低熔点琼脂糖凝胶纯化，与ｄＴ载体直接进行连接反应，再转化受体菌。运用该法对不同长度的病毒、细菌和动物细胞的ＤＮＡ片段进行了克隆试验，均获得高效率克隆。
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| 关 键 词： | 聚合酶链反应 克隆 PUC质粒 分子生物学 |
A METHOD OF EFFICIENTLY CLONING OF PCR PRODUCTS USING PUC PLASMID |
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| Ye Jun, Kong Jie, Liu Ping,Zhang Yan,Yang Conghai, Xu Huaishu.A METHOD OF EFFICIENTLY CLONING OF PCR PRODUCTS USING PUC PLASMID[J].Marine Fisheries Research,1995,16(1):72-75. |
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| Authors: | Ye Jun Kong Jie Liu Ping Zhang Yan Yang Conghai Xu Huaishu |
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| Abstract: | A method of efficiently cloning of PCR products using PUC plasmid was developed based on the template -independent terminal transferase activity of Taq polymerase.PUC plasmid was digested with Saml and incubated with Taq polymerase and dTTP to creat dT vector which has 3'thymidine overhangs. PCR products which have almost exclusively adenosine at the 3'end of fragment were gel purified and ligated to the vector without any enzymatic modification. The cloning procedure was proved to be simple,practical and efficient. |
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| Keywords: | PCR Cloning PUC plasmid dT vector |
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