What is the maximum length a dsDNA/duplex DNA can be synthesized?
The real answers lies in the resolution limit of the purification method and on the coupling efficiency of the DNA synthesizer. We can synthesize dsDNA/duplex DNA of 220 bases and obtain sufficient quantities by HPLC purification to perform successful gene construction. However, it should be remembered that the longer the DNA construct, the greater the chance of accumulated sequence errors.
Coupling efficiency is the factor that mainly affects the length of DNA that can be synthesized. Base composition and synthesis scales will also be contributing factors. The table below shows that at 99% coupling efficiency, a crude solution of synthesized 95-mers would contain 38% full-length product and 62% (n–x) failure sequences. This is without taking into account other chemical reactions, such as depurination (which mainly affects the base A). The frequency of depurination is small but increases significantly with the primer length. However, as we are very experienced in producing very long oligos, do not hesitate to discuss your projects with our specialists.