Highly accurate de novo polynucleotide synthesis
AssigneeTwist Bioscience Corporation
InventorsBill James Peck, Mary Noe, Stefan Pitsch, Patrick A. Weiss
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Abstract
Methods, systems, compositions, and devices for the manufacturing of high-quality building blocks, such as polynucleotides, are described herein. Processes described herein provide for efficient washing of residual reagents, solvents, or byproducts from previous synthetic steps to allow for the generation of polynucleotides with low error rates. Processes described herein also provide for reduction in deletion rates during chemical nucleic acid synthesis. Further, methods and devices described herein allow for the rapid construction and assembly of large libraries of highly accurate polynucleotides.
Claims22
A method of polynucleotide synthesis comprising:
- providing predetermined sequences for a plurality of preselected polynucleotides;
- providing a structure comprising a silicon surface;
- coupling at least one nucleoside to a polynucleotide attached to the silicon surface;
- washing the silicon surface with a wash solvent, wherein the wash solvent consists essentially of acetone;
- washing the silicon surface with a capping solution, the capping solution comprising:
- (i) at least one base;
- (ii) at least one O-nucleophile selected from a group consisting of acetic acid, formic acid, propionic acid, methoxyacetic acid, phenoxyacetic acid, and water;
- and
- (iii) tetrahydrofuran;
- oxidizing a linkage between the at least one nucleoside and the polynucleotide with an oxidizing solution;
- and
- repeating the coupling, washing, capping, and oxidizing steps to synthesize a plurality of polynucleotides.
A method for polynucleotide synthesis, comprising:
- providing predetermined sequences for a plurality of polynucleotides;
- providing a structure comprising a silicon surface, wherein the silicon surface comprises a plurality of discrete loci configured for polynucleotide synthesis;
- coupling at least one nucleoside to a polynucleotide attached to the silicon surface, according to the predetermined sequences;
- oxidizing a linkage between the at least one nucleoside and the polynucleotide with an oxidizing solution;
- and
- washing the silicon surface with a wash solvent, wherein the wash solvent consists essentially of acetone.
Dependencies are inferred from the claim text.