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Team:DTU-Denmark-2/Project/PlugnplayAssembly
From 2011.igem.org
Making Molecular Biology Easier
In 2009 the first team from DTU to participate in iGEM introduced the USER fusion Assembly standard. This year we introduce a more standardized version allowing easier use of the system.
We think that iGEM should be about assembling biobricks fast and combining them in any thinkable way with existing or new parts. Unfortunately classical cloning techniques can cause problems and even PCR can be cumbersome if you have little or no laboratory experience.
We therefore introduce a simple and fast way of building new devices or whatever you want with ready to use PCR products. All you have to do is to select your favorite bricks, mix them with the ready to use destination vector and 70 minutes later you are ready to transform your competent E. coli cells. No need to worry if your destination vector has been fully linearized, no need to perform site-directed mutagenesis to remove illegal restriction sites, just simple, fast, and easy.
Designing the system
When designing the system, the first thing we considered was which types of biobricks a device or other construct would consist of. We decided that the most systems would at least consist of a vector backbone, promoter, a gene of interest (GOI), a terminater, and a marker cassette.
For all these parts to be assembled by USER cloning it was important that the overhangs (tails) of the different parts were not identical. Designing a different tail for each end of a part was important to ensure directionality and correct order of the biobricks.
