Abstract
Over the past decades, remarkable progress on phosphoramidite chemistry-based large-scale de novo oligonucleotide synthesis has been achieved, enabling numerous novel and exciting applications. Among them, de novo genome synthesis and DNA data storage are striking. However, to make these two applications more practical, the synthesis length, speed, cost, and throughput require vast improvements, which is a challenge to be met by the phosphoramidite chemistry. Harnessing the power of enzymes, the recently emerged enzymatic methods provide a competitive route to overcome this challenge. In this review, we first summarize the status of large-scale oligonucleotide synthesis technologies including the basic methodology and large-scale synthesis approaches, with special focus on the emerging enzymatic methods. Afterward, we discuss the opportunities and challenges of large-scale oligonucleotide synthesis on de novo genome synthesis and DNA data storage respectively.
Highlights
De novo oligonucleotide synthesis is the synthesis of defined sequences of singlestrand nucleic acids
Despite the astonishing achievements on large-scale oligo synthesis that have been made in the past decades, our capacity to write DNA sequences still lags far behind our ability to read them
While the rapid development of DNA sequencing technology has heavily relied on the “sequencing by synthesis” method which can utilize the power of naturally formed polymerase for sequencing purpose, the oligo synthesis is dominated by chemical synthesis method with several limitations like limited synthesis length and environmentally hazardous, etc
Summary
De novo oligonucleotide synthesis (oligo synthesis) is the synthesis of defined sequences of singlestrand nucleic acids. Starting in the early 1990s, the microchipbased large-scale oligo synthesis methods were developed (Fodor et al, 1991; Pease et al, 1994; Singh-Gasson et al, 1999; Gao et al, 2001; Ghindilis et al, 2007; Ma et al, 2012; Kosuri and Church, 2014). Starting in the 1990s, oligo synthesis gradually developed into a higher throughput manner by parallel synthesis on a silica surface (Fodor et al, 1991; Pease et al, 1994) This microchipbased ( called ‘microarray-based’) large-scale oligo synthesis technologies, provides an inexpensive source of oligo building blocks for various applications (Ma et al, 2012; Kosuri and Church, 2014). The cost per nucleotide is between $0.00001 and 0.001 (Kosuri and Church, 2014)
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