|
|||||
|
|
| 有效扩增小片段DNA的一种方法 | |
| 引用本文： | 古先祥,李鸿岩,袁毅君.有效扩增小片段DNA的一种方法[J].天水师范学院学报,2011,31(2):29-32. |
| 作者姓名： | 古先祥 李鸿岩 袁毅君 |
| 作者单位： | 1. 复旦大学上海医学院细胞与遗传医学系,上海200433;张掖医学高等专科学校医学遗传学教研室,甘肃张掖734000 2. 复旦大学上海医学院细胞与遗传医学系,上海,200433 3. 张掖医学高等专科学校医学遗传学教研室,甘肃张掖,734000 |
| 摘 要： | 利用单链DNA连接酶可以连接单链DNA的性质将小片段DNA自连成环,随后利用phi29 DNA聚合酶进行恒温滚环复制,将扩增产物进行酶切,得到扩增后的小片段DNA.实验结果表明,单链DNA连接酶能有效地将30bp的小片段DNA自连成环,以phi29 DNA聚合酶扩增出大于10kb的DNA片段,产物经酶切后可进行第二轮扩增,并证明其成环方式为自成环.该方法能有效扩增小片段DNA. |
| 关 键 词： | 滚环复制 phi29 DNA聚合酶 小片段DNA |
A Method of Efficiency Amplification of Short DNA Fragments |
|
| Gu Xianxiang,Li Hongyan,Yuan Yijun.A Method of Efficiency Amplification of Short DNA Fragments[J].Journal of Tianshui Normal University,2011,31(2):29-32. | |
| Authors: | Gu Xianxiang Li Hongyan Yuan Yijun |
| Institution: | 1.Department of Cellular and Genetic Medicine,Shanghai Medical College,Fudan University,Shanghai 200433,China; 2.Department of Medical Genetics,Zhangye Medical College,Zhangye Gansu 734000,China; 3.School of Life Science and Chemistry,Tianshui Normal University,Tianshui Gansu 741001,China) |
| Abstract: | The aim is to establish a method of efficient amplification of small fragment DNA in this paper.The small DNA fragments were linked the ring by single-stranded DNA ligase(ssDNA ligase) which made single-strand DNA intramolecular ligation,and then carried out isothermal rolling circle replication by phi29 DNA polymerase.The amplified products were cut by incision enzyme and harvested the small fragment DNA.The results showed that ssDNA ligase made 30bp oligonucleotide DNA cyclization,and phi29 DNA polymerase amplified products more than 10kb in length.Amplification products were amplified second times after cut by Incision enzyme,and the way of single strand DNA cyclization was proved intramolecular.The method could be effectively applied to amplify the small DNA fragments. |
| Keywords: | rolling circle replication phi29 DNA polymerase small fragment DNA |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |