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Fig. 1 | Journal of Biological Engineering

Fig. 1

From: QuickStep-Cloning: a sequence-independent, ligation-free method for rapid construction of recombinant plasmids

Fig. 1

Overview of QuickStep-Cloning: a A schematic diagram presenting individual stages involved in the proposed method: (1) two parallel asymmetric PCRs of DNA of interest and PCR purification, (2) megaprimer-based PCR, (3) DpnI digestion, and (4) bacterial transformation. b Exemplary workflow for 1 kb insert and 7 kb cloning vector (exact duration of the asymmetric PCR depends on the length of cloned DNA fragment and the duration of megaprimer PCR is related to the size of the cloning vector). c Outline of exponential amplification taking place during QuickStep-Cloning – megaprimers anneal themselves to the product of linear amplification and are extended by polymerase, producing further copies of the two single-stranded templates in an exponential manner. It should be noted that for the given mechanism, exponential amplification occurs in parallel with the linear process

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