"
Materials and Methods
From 2008.igem.org
Materials and Methods
1. Cloning
Constructs were cloned by the AarI method. This a multi-part/combinatorial cloning method that is particularly well suited to shuffling protein domains with various promoters and terminators. The key to this approach is a Type II-S restriction enzyme. AarI is rare (7-cutter) that cuts at a site 4bp away from its binding site. Thus, AarI can generate four base overhangs of any sequence.
Furthermore, these overhangs are non-palindromic.
Digestion yields parts with standard, non-palindromic, overhangs.
AarI Shuttle Vector To facilitate exchange of parts between AarI users and the biobricks community, we are offering a Shuttle Vector. This vector accepts AarI parts in the AB or BD format (as were used in our 2008 project). These parts can then be cut out of the vector, with in-frame biobrick ends.
