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De novo synthesized gene libraries

  • US 9,833,761 B2
  • Filed: 10/16/2015
  • Issued: 12/05/2017
  • Est. Priority Date: 08/05/2013
  • Status: Active Grant
First Claim
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1. A method for synthesizing a library of oligonucleic acids encoding for predetermined sequences, the method comprising:

  • (a) providing predetermined sequences for a library of at least 100,000 non-identical oligonucleic acids, wherein the at least 100,000 non-identical oligonucleic acids collectively encode sequence for at least 750 genes;

    (b) providing a structure having a surface, wherein the surface comprises a plurality of clusters, wherein each cluster is selected for synthesis of oligonucleic acids associated with a single gene of the at least 750 genes, and wherein each cluster comprises a plurality of loci for oligonucleic acid extension;

    (c) adding a droplet of fluid comprising an extension reaction reagent specific to a locus, wherein the extension reaction reagent comprises a nucleoside phosphoramidite;

    (d) allowing sufficient time for an extension reaction step to occur;

    (e) repeating steps (c) and (d) to synthesize the at least 100,000 non-identical oligonucleic acids, wherein each non-identical oligonucleic acid is at least 30 bases in length and is attached to the surface; and

    (f) releasing the at least 100,000 non-identical oligonucleic acids from the surface, wherein the method comprises no amplification step prior to releasing the at least 100,000 non-identical oligonucleic acids from the surface.

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