The overall success of OE-PCR based DNA asseblies relies on several factors, being the most relevant ones the instrinsic features of the DNA sequence to assemble, the sequence and length of the overlapping overhangs, the design of outer primers for the final amplification and the conditions of the PCR reaction.
The efficacy of OE-PCR-based DNA assemblies depends on a number of crucial elements, including the intrinsic properties of the DNA sequence being assembled, the sequence and length of the complementary overhangs, the design of the outer primers for the final amplification step, and the conditions under which the PCR reaction is carried out.