Bridging PCR and partially overlapping primers for novel allergen gene cloning and expression insert decoration

Figure 1 Experimental design and steps involved in bridging PCR (BPCR) and partially overlapping primer-based PCR (POP-PCR). Gene fragments A and B had the junction region C. Under specific temperature conditions, the fragments would anneal together to form the template molecule, thus triggering the amplification cascade. The partially overlapping primers at each site of the template ends contained two primers pre-mixed in a certain molar ratio that had sequence identity but with different lengths. All the short primers located at the out-ward overhang region. The sequences of all primers used are shown in Table 1.