Abstract: A high-throughput automated gene synthesis device based on a cluster array includes a substrate and a microwell plate; the substrate is provided with a plurality of clusters of micropores; the inner wall surface of the micropores is chemically modified as a solid phase carrier for nucleic acid synthesis, or the micropore is filled with solid phase carriers; the clusters of micropores are arranged in a cluster array and each cluster of micropores has the same size and corresponding position as each well on the microwell plate. When using the device to synthesize oligonucleotides, by automatically recovering the synthesized oligonucleotides into a standard SBS plate of the corresponding size under the device, the oligonucleotide pool for each gene is formed. The yield of oligonucleotides is in picomole level, which is used for subsequent polymerase-mediated gene assembly (PCA) or ligase-mediated gene assembly (LCR) without amplification.

Abstract: A surface functionalizing method for use in high-throughput in situ synthesis of nucleic acids by 3D inkjet printing. The method includes subjecting a surface of a substrate to hydroxyl enrichment treatment; adding hydrophobic molecules to the surface of the substrate, the hydrophobic molecules being not reactive with phosphoramidite monomers; spraying, by a multi-channel piezoelectric inkjet head assembly, an etching ink to a predetermined area on the surface of the substrate for micro-etching, the etching ink being prepared with a fluoride compound reactive with the hydrophobic molecules; and adding hydrophilic molecules to the surface of the substrate. By using the method, a functionalized surface with given areas being patterned can be formed on the surface of the substrate, and then a same multi-channel piezoelectric inkjet head assembly can be directly used for subsequent high-resolution printing of phosphoramidite monomers and synthesis of nucleic acids.