Circular polymerase extension cloning of complex gene libraries and pathways.

High-throughput genomics and the emerging field of synthetic biology demand ever more convenient, economical, and efficient technologies to assemble and clone genes, gene libraries and synthetic pathways. Here, we describe the development of a novel and extremely simple cloning method, circular poly...

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Autores principales:	Jiayuan Quan, Jingdong Tian
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2009
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/92a3c3887a7948b7a29ab52222a24682
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spelling	oai:doaj.org-article:92a3c3887a7948b7a29ab52222a246822021-11-25T06:21:18ZCircular polymerase extension cloning of complex gene libraries and pathways.1932-620310.1371/journal.pone.0006441https://doaj.org/article/92a3c3887a7948b7a29ab52222a246822009-07-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19649325/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203High-throughput genomics and the emerging field of synthetic biology demand ever more convenient, economical, and efficient technologies to assemble and clone genes, gene libraries and synthetic pathways. Here, we describe the development of a novel and extremely simple cloning method, circular polymerase extension cloning (CPEC). This method uses a single polymerase to assemble and clone multiple inserts with any vector in a one-step reaction in vitro. No restriction digestion, ligation, or single-stranded homologous recombination is required. In this study, we elucidate the CPEC reaction mechanism and demonstrate its usage in demanding synthetic biology applications such as one-step assembly and cloning of complex combinatorial libraries and multi-component pathways.Jiayuan QuanJingdong TianPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 4, Iss 7, p e6441 (2009)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jiayuan Quan Jingdong Tian Circular polymerase extension cloning of complex gene libraries and pathways.
description	High-throughput genomics and the emerging field of synthetic biology demand ever more convenient, economical, and efficient technologies to assemble and clone genes, gene libraries and synthetic pathways. Here, we describe the development of a novel and extremely simple cloning method, circular polymerase extension cloning (CPEC). This method uses a single polymerase to assemble and clone multiple inserts with any vector in a one-step reaction in vitro. No restriction digestion, ligation, or single-stranded homologous recombination is required. In this study, we elucidate the CPEC reaction mechanism and demonstrate its usage in demanding synthetic biology applications such as one-step assembly and cloning of complex combinatorial libraries and multi-component pathways.
format	article
author	Jiayuan Quan Jingdong Tian
author_facet	Jiayuan Quan Jingdong Tian
author_sort	Jiayuan Quan
title	Circular polymerase extension cloning of complex gene libraries and pathways.
title_short	Circular polymerase extension cloning of complex gene libraries and pathways.
title_full	Circular polymerase extension cloning of complex gene libraries and pathways.
title_fullStr	Circular polymerase extension cloning of complex gene libraries and pathways.
title_full_unstemmed	Circular polymerase extension cloning of complex gene libraries and pathways.
title_sort	circular polymerase extension cloning of complex gene libraries and pathways.
publisher	Public Library of Science (PLoS)
publishDate	2009
url	https://doaj.org/article/92a3c3887a7948b7a29ab52222a24682
work_keys_str_mv	AT jiayuanquan circularpolymeraseextensioncloningofcomplexgenelibrariesandpathways AT jingdongtian circularpolymeraseextensioncloningofcomplexgenelibrariesandpathways
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Circular polymerase extension cloning of complex gene libraries and pathways.

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