Nature research paper: Continuous synthesis of E. coli genome sections and Mb-scale human DNA assembly
, In an odd step of CONEXER, recipient cells with a +1/−1 double selection cassette in their genome ; −1,) and a tetracycline resistance ) conferring plasmid encoding arabinose inducible lambda red components and Cas9 are mixed with donor cells and spotted on an agar plate. Donor cells contain an odd BAC and a non-transferable F’ plasmid. During incubation on the plate the BAC is conjugated from donor to recipient cells.
, In an even step of CONEXER, recipient cells with a +2/−2 double selection cassette in their genome ; −2, and a tetracycline resistance ) conferring plasmid encoding arabinose inducible lambda red components and Cas9 are mixed with donor cells and spotted on an agar plate. Donor cells contain an odd BAC and a non-transferable F’ plasmid. During incubation on the plate the BAC is conjugated from donor to recipient cells.
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