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Molecules 2012, 17(1), 328-340;

Enzyme Treatment-Free and Ligation-Independent Cloning Using Caged Primers in Polymerase Chain Reactions

Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro, Tokyo 153-8904, Japan
Department of Chemistry and Materials Engineering, Kansai University, 3-3-35 Yamate-cho, Suita, Osaka 564-8680, Japan
Authors to whom correspondence should be addressed.
Received: 28 October 2011 / Revised: 26 December 2011 / Accepted: 27 December 2011 / Published: 30 December 2011
(This article belongs to the Special Issue DNA-Templated Synthesis)
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A new simple scheme for constructing recombinant vectors that does not require any restriction enzyme, ligase, or any other special enzyme treatment has been developed. By using caged primers in PCR, unnatural sticky-ends of any sequence, which are sufficiently long for ligation-independent cloning (LIC), are directly prepared on the product after a brief UVA irradiation. Target genes and vectors amplified by this light-assisted cohesive-ending (LACE) PCR join together in the desired arrangement in a simple mixture of them, tightly enough to be repaired and ligated in competent cells. View Full-Text
Keywords: DNA; PCR; caged compounds; ligation; cloning DNA; PCR; caged compounds; ligation; cloning

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Kuzuya, A.; Tanaka, K.; Katada, H.; Komiyama, M. Enzyme Treatment-Free and Ligation-Independent Cloning Using Caged Primers in Polymerase Chain Reactions. Molecules 2012, 17, 328-340.

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