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Team:Cambridge/Gibson/Introduction
From 2010.igem.org
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Revision as of 00:18, 1 October 2010
Gibson Assembly: Introduction
Gibson Assembly is a technique for assembling DNA with short (c. 40 bp) overlapping sequences together. Since these overlapping regions can be easily added by PCR with primers which have added "flaps", any DNA sequences can be joined by this mechanism.
Advantages
- No scar created - useful for fusion proteins and adding an RBS, where scars can be problematic.
- Can re-ligate linear DNA into a circle - useful for site-directed mutagenesis
- Although it is roughly the same speed as standard BioBrick assembly for ligating two fragments, Gibson is perfect for doing multi part systems, since it takes the same amount of time to ligate n pieces of DNA together as it does for 2 pieces.
| Assembly Method | Number of steps to ligate N pieces of DNA |
| Standard Assembly | 
|
| Parallel Assembly | 
|
| Gibson Assembly | 
|
Disadvantages
- The need for planning, primers must be ordered in advance
- More expensive than BioBrick assembly
