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

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...

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Autores principales: Kuzuya, Akinori, Tanaka, Keita, Katada, Hitoshi, Komiyama, Makoto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2011
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6290560/
https://www.ncbi.nlm.nih.gov/pubmed/22210171
http://dx.doi.org/10.3390/molecules17010328
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author Kuzuya, Akinori
Tanaka, Keita
Katada, Hitoshi
Komiyama, Makoto
author_facet Kuzuya, Akinori
Tanaka, Keita
Katada, Hitoshi
Komiyama, Makoto
author_sort Kuzuya, Akinori
collection PubMed
description 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.
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spelling pubmed-62905602018-12-12 Enzyme Treatment-Free and Ligation-Independent Cloning Using Caged Primers in Polymerase Chain Reactions Kuzuya, Akinori Tanaka, Keita Katada, Hitoshi Komiyama, Makoto Molecules Article 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. MDPI 2011-12-30 /pmc/articles/PMC6290560/ /pubmed/22210171 http://dx.doi.org/10.3390/molecules17010328 Text en © 2012 by the authors; http://creativecommons.org/licenses/by/3.0/ licensee MDPI, Basel, Switzerland. This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Kuzuya, Akinori
Tanaka, Keita
Katada, Hitoshi
Komiyama, Makoto
Enzyme Treatment-Free and Ligation-Independent Cloning Using Caged Primers in Polymerase Chain Reactions
title Enzyme Treatment-Free and Ligation-Independent Cloning Using Caged Primers in Polymerase Chain Reactions
title_full Enzyme Treatment-Free and Ligation-Independent Cloning Using Caged Primers in Polymerase Chain Reactions
title_fullStr Enzyme Treatment-Free and Ligation-Independent Cloning Using Caged Primers in Polymerase Chain Reactions
title_full_unstemmed Enzyme Treatment-Free and Ligation-Independent Cloning Using Caged Primers in Polymerase Chain Reactions
title_short Enzyme Treatment-Free and Ligation-Independent Cloning Using Caged Primers in Polymerase Chain Reactions
title_sort enzyme treatment-free and ligation-independent cloning using caged primers in polymerase chain reactions
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6290560/
https://www.ncbi.nlm.nih.gov/pubmed/22210171
http://dx.doi.org/10.3390/molecules17010328
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AT katadahitoshi enzymetreatmentfreeandligationindependentcloningusingcagedprimersinpolymerasechainreactions
AT komiyamamakoto enzymetreatmentfreeandligationindependentcloningusingcagedprimersinpolymerasechainreactions

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