通过巢式PCR克隆ST13 cDNA的5'端片段
[Cloning the 5 ' end fragment of ST13 cDNA by nested PCR].
作者信息
Geng L, Cai X, Cao J, Fang Y
机构信息
Cancer Institute of Zhejiang Medical University, Hangzhou, 310009 P.R. China.
出版信息
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 1999 Jun;16(3):174-6.
OBJECTIVE
To clone and sequence the 5' -end fragment of ST13 cDNA.
METHODS
The 5' -end fragment of the ST13cDNA was amplified directly from cDNA library by nested PCR method, and cloned into pGEM-T. easy vector, then the sequencing of the inserted PCR product was performed.
RESULTS
After primary and secondary PCR, two PCR products obtained with the size of about 550bp and 480bp, respectively, were both proved to be the 5' -end of ST13 cDNA by sequencing.
CONCLUSION
the method presented is very simple and effective for cloning and sequencing the 5'-end of a target gene.
目的
克隆并测序ST13 cDNA的5′端片段。
方法
采用巢式PCR方法直接从cDNA文库中扩增ST13 cDNA的5′端片段，克隆至pGEM-T easy载体，然后对插入的PCR产物进行测序。
结果
经过一次和二次PCR，分别得到两条大小约为550bp和480bp的PCR产物，测序证明它们均为ST13 cDNA的5′端。
结论
所提出的方法对于目标基因5′端的克隆和测序非常简单有效。
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