Abstract
Vaccinia DNA topoisomerase I (TOPO) charged vectors with a sticky T are routinely used to clone polymerase chain reaction (PCR) products with an extra A at their 3′ end (TOPO TA Cloning from Invitrogen). TOPO charged blunt vectors are used to clone blunt end PCR products (TOPO Blunt Cloning). Here, we demonstrate that both TOPO TA vectors and TOPO Blunt vectors can be used to clone PCR products with either a blunt end or an extra A at the 3′ end. We further demonstrate that these vectors can be used to clone sticky end DNA generated with restriction enzymes. In summary, these TOPO vectors can be used as universal cloning vectors.
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References
Shuman, S. (1994) Novel approach to molecular cloning and polynucleotide synthesis using vaccinia DNA topoisomerase. J. Biol. Chem. 269, 32,678–32,684.
Mead, D. A., Pey, N. K., Herrnstadt, C., Marcil, R. A., Smith, L. M. (1991) A universal method for the direct cloning of PCR amplified nucleic acid. BioTechnology 9, 657–663.
Magnuson, V.L., Ally, D.S., Nylund, S.J., et al. (1996) Substrate nucleotide-determined non-templated addition of adenine by Taq DNA polymerase: implications for PCR-based genotyping and cloning. Biotechniques 21, 700–709.
Cheng, C. H. and Shuman, S. (2000) DNA strand transfer catalyzed by vaccinia topoisomerase ligation DNAs containing a 3′ mononucleotide overhang. Nucleic Acids Res. 28, 1893–1898.
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Geng, L., Xin, W., Huang, DW. et al. A universal cloning vector using vaccinia topoisomerase I. Mol Biotechnol 33, 23–28 (2006). https://doi.org/10.1385/MB:33:1:23
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DOI: https://doi.org/10.1385/MB:33:1:23