Fig. 3

Investigation and optimization of asymmetric PCR stage of QuickStep-Cloning. a Yield of two parallel asymmetric PCRs for different primer ratios (represented by two separate rows for each ratio). b Identification of single stranded product of asymmetric PCR stage – (1) individual products of two parallel asymmetric PCRs, (2) both products after 2 min denaturation at 94 °C, (3) renatured products, and (4) products of two parallel asymmetric PCRs after being mixed together. In all gel pictures, the appearance of low and high molecular weight bands could be attributed to non-specific binding of primers, commonly seen in regular PCRs