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[Cloning the 5 ' end fragment of ST13 cDNA by nested PCR]
1Cancer Institute of Zhejiang Medical University, Hangzhou, 310009 P.R. China. jcao@public.hz.zj.cn
Abstract
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.