OEPR Cloning: an Efficient and Seamless Cloning Strategy for Large- and Multi-Fragments

Here, an efficient cloning strategy for large DNA fragments and for simultaneous assembly of multiple DNA fragments assembly is presented. This strategy is named OEPR (based on O verlap E xtension P CR and R ecombination in vivo ). OEPR cloning is a seamless, restriction- and ligation-independent me...

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Bibliographic Details
Published in:Scientific reports 2017-03, Vol.7 (1), p.44648-44648, Article 44648
Main Authors: Liu, Chang-Jun, Jiang, Hui, Wu, Lei, Zhu, Ling-Yun, Meng, Er, Zhang, Dong-Yi
Format: Article
Language:English
Subjects:
631/1647/1513/2216
631/337/149
Animals
Cloning
Cloning, Molecular - methods
Deoxyribonucleic acid
DNA
DNA - genetics
E coli
Enzymes
Homologous recombination
Homologous Recombination - genetics
Humanities and Social Sciences
Humans
Methods
multidisciplinary
Mutagenesis, Insertional - genetics
Polymerase chain reaction
Polymerase Chain Reaction - methods
Rats
Science
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Summary:Here, an efficient cloning strategy for large DNA fragments and for simultaneous assembly of multiple DNA fragments assembly is presented. This strategy is named OEPR (based on O verlap E xtension P CR and R ecombination in vivo ). OEPR cloning is a seamless, restriction- and ligation-independent method. The method takes advantage of both homologous recombination enzymes in E. coli and overlap PCR. Using OEPR cloning, a long fragment (1–6 kb) or multiple fragments (2–4 fragments) can be easily constructed and simultaneously assembled into a target vector.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep44648