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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Main Authors: Jiayuan Quan, Jingdong Tian
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2009-07-01
Series:PLoS ONE
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19649325/pdf/?tool=EBI
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author Jiayuan Quan
Jingdong Tian
author_facet Jiayuan Quan
Jingdong Tian
author_sort Jiayuan Quan
collection DOAJ
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.
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spelling doaj.art-92a3c3887a7948b7a29ab52222a246822022-12-21T21:53:00ZengPublic Library of Science (PLoS)PLoS ONE1932-62032009-07-0147e644110.1371/journal.pone.0006441Circular polymerase extension cloning of complex gene libraries and pathways.Jiayuan QuanJingdong TianHigh-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.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19649325/pdf/?tool=EBI
spellingShingle Jiayuan Quan
Jingdong Tian
Circular polymerase extension cloning of complex gene libraries and pathways.
PLoS ONE
title 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_short Circular polymerase extension cloning of complex gene libraries and pathways.
title_sort circular polymerase extension cloning of complex gene libraries and pathways
url https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19649325/pdf/?tool=EBI
work_keys_str_mv AT jiayuanquan circularpolymeraseextensioncloningofcomplexgenelibrariesandpathways
AT jingdongtian circularpolymeraseextensioncloningofcomplexgenelibrariesandpathways