Self-Assembly Cloning: A Rapid Construction Method for Recombinant Molecules from Multiple Fragments | Zendy

Akira Matsumoto | Zendy; Taichi Q. Itoh | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

Self-Assembly Cloning: A Rapid Construction Method for Recombinant Molecules from Multiple Fragments

Author(s) -

Akira Matsumoto,

Taichi Q. Itoh

Publication year - 2011

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/000113705

Subject(s) - recombinant dna , cloning (programming) , plasmid , computational biology , fusion protein , fragment (logic) , molecular cloning , fusion , dna , domain (mathematical analysis) , base pair , fusion gene , microbiology and biotechnology , biology , gene , chemistry , genetics , algorithm , computer science , mathematics , peptide sequence , linguistics , philosophy , programming language , mathematical analysis

Enzyme-free cloning (EFC) can rapidly produce an in-frame fusion gene with multiple fragments. To practically apply EFC, we investigated the extent and sequence of complementary staggered overhangs necessary to direct self-assembly of multiple fragments as well as a size limitation of the constructed DNA molecule. Six-base pair overhangs with 50% GC content were sufficient to direct self-assembly. A functional plasmid that exceeded 10 kb, which includes an in-frame fusion domain, was efficiently constructed from four PCR fragments in one step by our improved method.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research