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. 2011 Jun 2;6(6):e20556. doi: 10.1371/journal.pone.0020556

Figure 10. Construction of QC cloning vectors.

Figure 10

QC cloning vectors can be prepared by amplification of a DNA fragment containing a visible selectable marker (a lacZα fragment was used here) with two primers with 5′ extensions containing the bap2 sequence (blue box) and the catching sequence (CS, red box). The primers also contain extensions C and D with homology with any cloning vector of choice. The PCR product is cloned by ligation-independent cloning in a linearized vector (here pICH36833) with DNA ends homologous to sequences C and D. DNA from blue colonies are sequenced to make sure that no mutations are present in the sequence of bap2 and the CS.