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Team:Lethbridge CCS/Project
From 2008.igem.org
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While there is an established BioBrick format, there is not yet a standard method for turning a gene of interest into a BioBrick. | 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 | 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.' | + | 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 [http://nar.oxfordjournals.org/cgi/content/abstract/18/20/6069 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.' |
=== '''Rationale''' === | === '''Rationale''' === | ||
Revision as of 21:12, 28 October 2008
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 [http://nar.oxfordjournals.org/cgi/content/abstract/18/20/6069 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.'
Rationale
The underlying idea of iGEM is that an engineering approach can be applied to biology. Using a computer analogy, we would like to have software (DNA sequences) that can be run on compatible hardware (microorganisms) anywhere in the world. Ideally, this software-hardware interface would be highly portable, but also simple. The BioBricks used by iGEM are advertised with 'Lego' blocks, implying a very easy-to-use set of tools. Our team of high school students discovered that (at this stage, at least) synthetic biology is NOT yet at the stage of quick and easy implementation in just anyone's kitchen or garage.
Plasmid: [http://partsregistry.org/Part:BBa_K155000 K155000]
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