| 摘要 |
Provided is a method of mass-producing high-purity nucleotides including providing a sequencing substrate having a clonal library of oligonucleotides on a solid support, sequencing the clonal library, obtaining measured location data of the solid support on the sequencing substrate, mapping pixel data of a signal generated from the solid support given as a result of the sequencing with the measured location data, extracting the solid support having a desired base sequence from the sequencing substrate using the mapping result, and amplifying an oligonucleotide on the extracted solid support to replicate on a large scale. |
| 主权项 |
1. A method of producing high-purity oligonucleotides, comprising the steps of:
providing a sequencing substrate having a clonal library of oligonucleotides on solid supports, wherein the solid supports are disposed in a predetermined section of the sequencing substrate; sequencing the clonal library and generating optical or electromagnetic signals of the clonal library; obtaining pixel data of a signal generated from each of the solid supports having a selected DNA sequence during the sequencing step; obtaining measured location data of the solid support on the substrate by scanning the substrate with an imaging device; mapping the pixel data with the measured location data of the solid supports, wherein an algorithm is used in the step of mapping for tracing an exact location of each solid support; performing the algorithm comprising: dividing an entire domain of the substrate into a plurality of spatially small subdomains,
selecting one subdomain (i, j) among the plurality of subdomains, the one subdomain defines where a location of the pixel data and the measured location data is matched,defining a transform function of the one subdomain by applying the matched pixel data and the measured location data,defining an input queue having adjacent subdomains which are adjacent to the one subdomain (i, j), wherein the adjacent subdomains are (i−1, j−1), (i−1, j), (i−1, j+1), (i, j−1), (i, j+1), (i+1, j−1), (i+1, j), and (i+1, j+1),defining a next transform function by applying any one of the adjacent subdomains to the transform function which is previously defined,wherein the applied adjacent subdomain is added into a complete queue so that the applied adjacent subdomain is not repeatedly applied, wherein the complete queue includes the adjacent subdomains different from the input queue,wherein an order of applying the adjacent subdomain defines a matching direction, anddetermining transform functions corresponding to each subdomain of the entire domain by expanding the matching direction via repeating the step of defining the input queue and the stet of defining the next transform function, wherein at least three mapped data are provided to trace the exact location of each solid support; extracting the solid supports having the selected DNA sequence from the sequencing substrate by a pulsed laser beam, wherein each solid support is extracted based on the algorithm; and amplifying an oligonucleotide on the extracted solid supports to produce a plurality of the high-purity oligonucleotides by replication. |
