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APPLICATION RESEARCH OF TA CLONING

Yu Xiu

Open publisher page 1 citations

Abstract

Objective:To study a rapid and high efficiency cloning technique of PCR products.Methods:To prepare T vector,the pUC18 plasmid was digested to complation with SmaI,and the liner pUC18 was tailed with Taq DNA polymerase and tTTP for 2h at 75℃.PCR products of E6 fragments amplified from plasmid pBR322 HPV16 were ligated into the T tailed pUC18 vector.The recombinant clones were confirmed by PCR and restriction endonuclease cleaving analysis.Results:The recombinant plasmid pUC18 E6 had been got.Conclusion:TA cloning plus low temperature long time link is a highly efficient and rapid cloning technique of PCR products.

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What this paper is about

Objective:To study a rapid and high efficiency cloning technique of PCR products.Methods:To prepare T vector,the pUC18 plasmid was digested to complation with SmaI,and the liner pUC18 was tailed with Taq DNA polymerase and tTTP for 2h at 75℃.PCR products of E6 fragments amplified from plasmid pBR322 HPV16 were ligated into the T tailed pUC18 vector.The recombinant clones were confirmed by PCR and restriction endonuclease cleaving analysis.Results:The recombinant plasmid pUC18 E6 had been got.Conclusion:TA cloning plus low temperature long time link is a highly efficient and rapid cloning technique of PCR products.

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Available abstract

Objective:To study a rapid and high efficiency cloning technique of PCR products.Methods:To prepare T vector,the pUC18 plasmid was digested to complation with SmaI,and the liner pUC18 was tailed with Taq DNA polymerase and tTTP for 2h at 75℃.PCR products of E6 fragments amplified from plasmid pBR322 HPV16 were ligated into the T tailed pUC18 vector.The recombinant clones were confirmed by PCR and restriction endonuclease cleaving analysis.Results:The recombinant plasmid pUC18 E6 had been got.Conclusion:TA cloning plus low temperature long time link is a highly efficient and rapid cloning technique of PCR products.

Key concepts: Cloning (programming), Cloning vector, Plasmid, Restriction enzyme, Recombinant DNA, Molecular biology, Multiple cloning site, Molecular cloning

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