US 11,691,118 B2
Devices and methods for oligonucleic acid library synthesis
William Banyai, San Francisco, CA (US); Bill James Peck, Santa Clara, CA (US); Andres Fernandez, San Francisco, CA (US); Siyuan Chen, San Mateo, CA (US); Pierre Indermuhle, Berkeley, CA (US); and Eugene P. Marsh, El Granada, CA (US)
Assigned to TWIST BIOSCIENCE CORPORATION, South San Francisco, CA (US)
Filed by Twist Bioscience Corporation, South San Francisco, CA (US)
Filed on Jul. 6, 2020, as Appl. No. 16/921,712.
Application 16/921,712 is a continuation of application No. 15/960,319, filed on Apr. 23, 2018, granted, now 10,744,477.
Application 15/960,319 is a continuation of application No. 15/135,434, filed on Apr. 21, 2016, granted, now 9,981,239, issued on May 29, 2018.
Claims priority of provisional application 62/220,856, filed on Sep. 18, 2015.
Claims priority of provisional application 62/150,795, filed on Apr. 21, 2015.
Prior Publication US 2020/0330953 A1, Oct. 22, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. B01J 19/00 (2006.01); C12N 15/10 (2006.01); B01L 3/00 (2006.01)
CPC B01J 19/0046 (2013.01) [C12N 15/1093 (2013.01); B01J 2219/00317 (2013.01); B01J 2219/00497 (2013.01); B01J 2219/00511 (2013.01); B01J 2219/00522 (2013.01); B01J 2219/00585 (2013.01); B01J 2219/00596 (2013.01); B01J 2219/00635 (2013.01); B01J 2219/00637 (2013.01); B01J 2219/00662 (2013.01); B01J 2219/00722 (2013.01); B01L 3/50857 (2013.01)] 15 Claims
 
1. A method for de novo oligonucleotide synthesis, comprising:
(a) providing predetermined sequences for a plurality of non-identical oligonucleotides;
(b) providing a device for synthesizing oligonucleotides, comprising:
(i) a solid substrate;
(ii) a main channel, wherein the main channel extends vertically into the solid substrate from an opening on a top side of the solid substrate; and
(iii) a plurality of microchannels, wherein each microchannel of the plurality of microchannels extends vertically from an opening on a bottom side of the solid substrate into the main channel, and wherein each microchannel of the plurality of microchannels comprises a fluid path having at least one turn of 45 degrees to 180 degrees in total, when viewed from a top view, and
wherein the device comprises more than 20,000 of the microchannels in total;
(c) adding a droplet of fluid comprising an extension reaction reagent specific to a microchannel;
(d) allowing sufficient time for an extension reaction step to occur; and
(e) repeating steps (c) and (d) until the plurality of non-identical oligonucleotides are synthesized.