Construction of QC cloning vectors.

Abstract

<p>QC cloning vectors can be prepared by amplification of a DNA fragment containing a visible selectable marker (a <i>lacZ</i>α 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.</p