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Team:Lethbridge CCS/Project
From 2008.igem.org
Our project centers on the development and optimization of a ligase-independent cloning (LIC) protocol.
Abstract:
Ligase-Independent Cloning as a Standard for BioBrick Preparation
While there is an established BioBrick format, there is not yet a standard method for turning a gene of interest into a BioBrick. Ideally, such a standard method would be easily adopted, even by amateurs, and would lend itself to automation. A significant drawback of several existing techniques is their dependence on ligase treatment, which is often problematic. We propose a ligase-independent cloning (LIC) method, based on the technique of Aslanidis & de Jong (1990), as a possible standard for novel BioBrick preparation. Instead of short overhangs and ligase treatment, LIC uses long overhangs to circularize plasmid vectors for transformation without the use of ligase. The LIC method reduces the number of enzyme steps required for cloning, thus lending itself to easy adoption, automation, and real biological 'engineering.'
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