US 12,226,766 B2
Removing and reinserting protein nanopores in a membrane using osmotic imbalance
Geoffrey Barrall, San Diego, CA (US); Takeshi Harada, San Jose, CA (US); Jason Komadina, Fremont, CA (US); Pirooz Parvarandeh, Los Altos Hills, CA (US); and Charlotte Yang, San Jose, CA (US)
Assigned to Roche Sequencing Solutions, Inc., Pleasanton, CA (US)
Filed by Roche Sequencing Solutions, Inc., Pleasanton, CA (US)
Filed on Apr. 20, 2020, as Appl. No. 16/853,624.
Application 16/853,624 is a continuation of application No. PCT/EP2018/078925, filed on Oct. 22, 2018.
Claims priority of provisional application 62/575,814, filed on Oct. 23, 2017.
Prior Publication US 2020/0246791 A1, Aug. 6, 2020
Int. Cl. B01L 3/00 (2006.01); C12Q 1/6869 (2018.01)
CPC B01L 3/502 (2013.01) [C12Q 1/6869 (2013.01); B01L 2200/12 (2013.01); B01L 2200/143 (2013.01); B01L 2300/044 (2013.01); B01L 2300/0663 (2013.01); B01L 2300/0896 (2013.01); B01L 2400/06 (2013.01)] 9 Claims
OG exemplary drawing
 
1. A system for updating a sequencing cell, the system comprising:
a fluid chamber comprising a first electrolyte reservoir of the sequencing cell, a second electrolyte reservoir of the sequencing cell, and a membrane that divides the first electrolyte reservoir from the second electrolyte reservoir, wherein the membrane comprises an initial nanopore, and wherein the first electrolyte reservoir has a first initial osmolarity and the second electrolyte reservoir has a second initial osmolarity;
a flow subsystem that controls a flow of a solution to the first electrolyte reservoir;
a plurality of replacement nanopores;
a voltage source configured to apply a voltage across the membrane; and
a processor communicably coupled with the voltage source and the flow subsystem and configured to perform operations comprising:
flowing a first electrolyte solution to the first electrolyte reservoir, wherein the first electrolyte solution has a first electrolyte solution osmolarity that is different than the first initial reservoir osmolarity, thereby causing the first electrolyte reservoir to have a new osmolarity that is different than the first initial osmolarity, so as to eject the initial nanopore from the membrane;
adding a second electrolyte solution to the first electrolyte reservoir, wherein the second electrolyte solution comprises the plurality of replacement nanopores, and wherein the second electrolyte solution has a second electrolyte solution osmolarity that is closer to the second initial osmolarity than the first electrolyte solution osmolarity; and
applying a voltage across the membrane so as to insert one of the plurality of replacement nanopores into the membrane.