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Rapid hierarchical assembly of medium-size DNA cassettes.

ABSTRACT: Synthetic biology applications call for efficient methods to generate large gene cassettes that encode complex gene circuits in order to avoid simultaneous delivery of multiple plasmids encoding individual genes. Multiple methods have been proposed to achieve this goal. Here, we describe a novel protocol that allows one-step cloning of up to four gene-size DNA fragments, followed by a second assembly of these concatenated sequences into large circular DNA. The protocols described here comprise a simple, cheap and fast solution for routine construction of cassettes with up to 10 gene-size components.

SUBMITTER: Schmid-Burgk JL

PROVIDER: S-EPMC3384347 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature

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Rapid hierarchical assembly of medium-size DNA cassettes.

Schmid-Burgk Jonathan Leo JL Xie Zhen Z Frank Stefan S Virreira Winter Sebastian S Mitschka Sibylle S Kolanus Waldemar W Murray Andrew A Benenson Yaakov Y

Nucleic acids research 20120315 12

Synthetic biology applications call for efficient methods to generate large gene cassettes that encode complex gene circuits in order to avoid simultaneous delivery of multiple plasmids encoding individual genes. Multiple methods have been proposed to achieve this goal. Here, we describe a novel protocol that allows one-step cloning of up to four gene-size DNA fragments, followed by a second assembly of these concatenated sequences into large circular DNA. The protocols described here comprise a ...[more]

PMID: 22422837

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