Barrick Lab :: ProtocolsCIDARMoclo

---+ Modular Cloning using CIDAR MoClo kit

The lab recently acquired a CIDAR MoClo (_C_ ross-disciplinary _I_ ntegration of _D_ esign _A_ utomation _R_ esearch lab _Mo_ dular _Clo_ ning) Kit from Addgene.  The website describing its use can be found here: [[https://www.addgene.org/cloning/moclo/densmore/][Addgene page describing CIDAR MoClo]].

The original paper can be found here: [[http://www.ncbi.nlm.nih.gov/pubmed/26479688][Paper describing updated CIGAR MoClo]]

In short, MoClo is a golden-gate based assembly strategy that allows for combinatorial assembly of plasmids based on individual parts.  So if you have a particular gene you want to express (say gfp), and want to try a variety of promoters and ribosome binding sites, this is a great option for easily making these variants (90 minute assembly).

However- all of your parts must be on "entry vectors."  This process can be a little time consuming and requires sequence verification.  So if you're only going to build a single construct, and aren't interested in trying a lot of variants, Gibson may make more sense.  

 


-- Main.SeanLeonard - 03 Feb 2016

Barrick Lab > ProtocolList > ProtocolsCIDARMoclo

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SeanLeonard

Topic revision: r1 - 2016-02-03 - 22:25:48 - Main.SeanLeonard