Workflow diagram of clone production. The entire process from the design of primers to production of clone stocks is shown for the four production phases. The process started by designing primers for every ORF in the genome. The primers were used to amplify the ORFs from the genome. Subsequent amplifications with universal primers generated ORF sequences flagged by recombinational cloning sites at either end and were monitored by a diagnostic gel. The product was cloned into a recombinational cloning vector via a BP clonase reaction or In-Fusion reaction. Competent bacterial strains were transformed with the reaction mix to yield colonies that were isolated robotically, cultured in liquid medium, and stored as 15% glycerol stocks.
