Functionalized surface treatment method for 3D ink jet method high-flux nucleic acid in-situ synthesis

Abstract

The invention discloses a functionalized surface treatment method for high-flux nucleic acid in-situ synthesis through a 3D ink jet method. The method comprises the following steps: carrying out hydroxylation enrichment treatment on the surface of a substrate; modifying hydrophobic molecules on the surface of the substrate; the hydrophobic molecule does not react with the phosphoramidite monomer; spraying etching ink to a preset area on the surface of the substrate by adopting a multi-channel piezoelectric ink-jet head assembly for micro-etching; the etching ink is prepared from fluorine salt compounds subjected to hydrophobic molecular reaction; and modifying hydrophilic molecules on the surface of the substrate. According to the method disclosed by the invention, a functional surface with a patterned specific area can be formed on the surface of the substrate, then subsequent high-resolution phosphoramidite monomer and nucleic acid synthesis can be directly carried out by using the same multi-channel piezoelectric ink jet head assembly, and compared with the existing method, expensive photoetching equipment and repeated position calibration design are not needed, so that the synthesis cost is reduced; the density of available active sites on the functionalized sites can also be improved, and the yield of single oligonucleotide is improved.

Description

technical field [0001] The invention relates to a functionalized surface treatment method for high-throughput nucleic acid in-situ synthesis by 3D inkjet method, which belongs to the field of synthetic biology. Background technique [0002] High-throughput oligonucleotide synthesis as a powerful tool in synthetic biology. The traditional oligonucleotide synthesis method is column synthesis based on phosphoramidite chemistry, each oligonucleotide is synthesized in a separate synthesis tube or in a column in a synthesis plate, and the yield of each nucleic acid is high , generally in the order of nmol, but the amount of reagents consumed in the synthesis is high and the cost is high, and the synthesis throughput is limited. The new generation of DNA synthesis technology mainly realizes the high-throughput synthesis of oligonucleotides through massively parallel synthesis technology. High-throughput oligonucleotide synthesis mainly includes photochemical chip in situ synthesi...

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Problems solved by technology

[0004] The purpose of the present invention is to provide a functionalized surface treatment method for 3D inkjet method high-throughput nucleic acid in situ synthesis, the surface treated by the present invention A multi-channel piezo inkjet head assembly can be used for simultaneous droplet in situ etching and subsequent addition of functional groups to the surface for efficient calibration-free oligo synthesis, addressing functions that currently require the use of complex lithographic techniques and alignment devices The high cost of the chemical surface treatment method and the contradiction that the output and throughput of the traditional glass slide functional surface treatment method cannot be improved at the same time

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