An oligonucleotide synthesizer based on a microreactor chip and an inkjet printer

Abstract

Synthetic oligonucleotides (oligos) are important tools in the fields of molecular biology and genetic engineering. For applications requiring a large number of oligos with high concentration, it is critical to perform high throughput oligo synthesis and achieve high yield of each oligo. This study reports a microreactor chip for oligo synthesis. By incorporating silica beads in the microreactors, the surface area of the solid substrate for oligo synthesis increases significantly in each microreactor. These beads are fixed in the microreactors to withstand the flushing step in oligo synthesis. Compared to conventional synthesis methods, this design is able to avoid protocols to hold the beads and integrate more microreactors on a chip. An inkjet printer is utilized to deliver chemical reagents in the microreactors. To evaluate the feasibility of oligo synthesis using this proof-of-concept synthesizer, an oligo with six nucleotide units is successfully synthesized.

Inkjet Bioprinting of Biomaterials

“…(c) The four-step for oligonucleotide synthesis on the silica beads: deprotection, coupling, capping, and oxidation. Reproduced with permission from ref . Copyright 2019 Springer Nature under a CC BY 4.0 License .…”

“…Because of the noncontact characteristic during printing, the surface of substrate has no influence on the printer so that substrate can be modified to enhance the yield of synthesis. Li et al increased the surface area of a microreactor via adding silica beads to each microreactor point . As shown in Figure B, the chemical reagents were delivered into the microreactor via inkjet printing according to the pre-established oligo sequence for synthesis.…”

“…Li et al increased the surface area of a microreactor via adding silica beads to each microreactor point. 197 As shown in Figure 16B, the chemical reagents were delivered into the microreactor via inkjet printing according to the pre-established oligo sequence for synthesis. Surface area used for oligonucleotide synthesis increased by 71 times with the existence of the silica beads.…”

The evolving art of creating genetic diversity: From directed evolution to synthetic biology

“…Technological advances to miniaturise phosphoramidite chemistry using solid-phase supported synthesis ( LeProust et al, 2001 ; LeProust et al, 2010 ; Tian et al, 2004 ) utilise inkjet ‘printing’ techniques to deliver reagents for polymerisation on a miniscule scale. This technology reduces synthesis volumes by >10 6 (to sub-femtomole scale ( Li et al, 2019 )) and thus permits thousands of individual oligonucleotides sequences to be synthesised in parallel at low cost. Consequently, each individual variant sequence of a library can be specifically synthesised in parallel, which when cleaved from the solid support creates a pool of oligonucleotides containing only the desired variant sequences ( Fig.…”

Microfluidic Pneumatic Printed Sandwiched Microdroplet Array for High-Throughput Enzymatic Reaction and Screening

“…19,20 Nowadays, it has begun attracting people's attention to replace traditional inks with chemical or biological reagents. [21][22][23][24][25] Such methods have been applied to many biochemical fields, including combinatorial synthesis, [26][27][28] drug screening, 29,30 chemical dilution, 31 and gene analysis. 32 The advantages of inkjet printing include its high-throughput, flexibility and high resolution.…”