From 2011.igem.org
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The use of restriction digestion and ligation although developed over 20 years ago, remains to be the standard cloning technique. The use of restriction enzymes can be both a time consuming and cumbersome process. To optimize the cloning process to become faster and more efficient, it is desirable to avoid the use of restriction enzymes.
Back in 2003 the USER friendly cloning was developed by New England Biolabs to ensure cloning without the use of restriction enzymes. Further development of the method in recent years has meant that construction of the vectors used for USER cloning can now be made quickly and efficiently without the use of restriction enzymes.
We are developing a standardized system, where certain categories of biological parts can be gathered. The idea is that the parts are in microtiter plates where they can be directly mixed with other desired parts and an expression vector and then can be assembled within a few hours. Hereby purification of digested vectors and PCR-products and ligation are avoided. To show how easy it is, we will demonstrate the use of this system by developing a reporter targeting system for the fungus Aspergillus niger and furthermore demonstrate its application in mammalian cells.
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