ABSTRACT
Several alternative methods for gene synthesis have been developed over the years including gene synthesis on solid phase in several related formats [11-14], FokI based synthesis [15], polymerase cycling assembly [16,17], ligase chain reaction [18], shotgun synthesis [19], synthesis by serial cloning [20], and insertional gene synthesis [21]. e many methods available each have their distinctive advantages and disadvantages. Some methods are better for GC rich sequences, some for highly repetitive sequences. Some methods are faster and some have higher accuracy. ere is no “best method”, but instead the many methods complement each other giving the user a plethora of gene synthesis tools to choose from. With the appearance of several commercial gene synthesis vendors, such as DNA2.0, most scientists can now quickly and routinely order any synthetic DNA fragment without having to ponder the alternative synthesis methods. With the many rapidly advancing gene synthesis technologies available, essentially any sequence can be synthesized independent of sequence characteristics and requirements.
