35-Fragment Golden Gate Assembly with BsmBI-v2*

(A) Schematic of the 35-fragment lac operon cassette test system. (B) Results of the assembly reactions. Four replicate experiments were carried out to quantify the number of colony-forming units harboring correct and incorrect assembly products per 50 µl of E. coli outgrowth plated (0.1 µl of the assembly reaction). On average, 71% of the observed transformants harbored correctly assembled products.  (C) Representative agar plate with blue and white colonies. Blue transformants harbor correct assembly constructs, and white transformants harbor inaccurate assembly products. For more information, please see reference 2.



* Complexities up to 24 fragments have been routinely achieved with both precloned and amplicon insert test systems.  Complexities of 35+ fragments have only used amplicon inserts to date,  as we have not yet constructed a 35 fragment precloned insert test system.