摘要
Ｂｌｕｅｓｃｒｉｐｔ经ＥｃｏＲＶ酶切后，在Ｔａｑ酶作用下加入ｄＴＴＰ使其３端加上碱基“Ｔ”而成为３端突出的粘性末端载体。用此载体直接与ＰＣＲ产物连接进行克隆。
Bluescript plasmid is used to reconstruct a T tail vector for direct cloning of PCR products. The plasmids containing the site of EcoR V are digested with EcoR V and incubated with Taq polymerase under such condition: 1 U Taq polymerase, 1μg plasmids, 2 mmol/L dTTP, 20 μl column in 70℃ for 2 hours. After phenol extraction and precipitation,the T vector is ready for cloning. PCR products of HCV,dengue virus genes IFN and IL 2 gene are ligated with the T vector directly,and cloned.The result shows that this procedure with T vector is about 50 to 100 fold more efficient than attempting to clone the PCR products into a blunt ended cut vector.
出处
《军事医学科学院院刊》
CSCD
北大核心
1996年第4期293-295,共3页
Bulletin of the Academy of Military Medical Sciences
关键词
聚合酶链反应
载体
克隆
polymerase chain reaction
vector
clone
