Figure 8
From: ZeBRα a universal, multi-fragment DNA-assembly-system with minimal hands-on time requirement
Effective, high fidelity five-fragment ZeBRα-assembly with OTG derived autoinduced PPY-extracts. (a) ZeBRα assembly of four fragments into pT7-HindIII-ccdB results in the triple-antibiotic resistant pZeBRα5 for convenient screening of successful recombinants. (b) The resistance against ampicillin and tetracycline critically depends on in-frame fusion of the respective ORF and serves therefore as estimate for the error-rates of the recombinogenic extracts. (c) Colonies displaying the double and triple antibiotic phenotypes expressed as cfu/µg transformed DNA expected for PCR-template carryover (pBR322) and the pZeBRa5 (dashed line). (d) Gel image of RsaI digested plasmids prepared from triple resistant colonies and the pBR322-vector that was used to generate the PCR-fragments for the assemblies.
