New method for synthesizing polynucleotides using various libraries of oligonucleotides

摘要

A method for synthesizing a target double-stranded (ds) polynucleotide having a predetermined sequence, a) providing an oligonucleotide library comprising a diversity of oligonucleotide library members in an array device, each of the library members having a different nucleotide sequence and contained in an individual library containment in aqueous solution, the diversity being at least one protrusion A step comprising a single-chain oligonucleotide having (ss oligo) and double-stranded oligonucleotide (ds oligo) and covering at least 10,000 pairs of matched oligonucleotides, b) In a first step, using a liquid handler to transfer at least a first pair of matched oligonucleotides from the library to a first reaction container, assemble the matched oligonucleotides, to a first reaction comprising at least one protrusion Obtaining a product, c) In the second and further steps, using a liquid handler, transfer at least a second and additional pair of matched oligonucleotides from the library to the second and further reaction containers, respectively, and assemble the matched oligonucleotides to at least each. Obtaining second and further reaction products, each comprising one protrusion, d) by assembling the first, second and further reaction products into a predetermined workflow, thereby generating the target ds polynucleotide having a protrusion, and optionally performing a finalization step to produce a smooth end, The pair of matched oligonucleotides used in steps b) and c) and the assembly workflow are determined using an algorithm that generates the target ds polynucleotide.