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US 9,850,534 B2Granted
Methods for creating bilayers for use with nanopore sensors
OwnerRandall Davis, Roger Cheninventors
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Abstract
The present disclosure provides biochips and methods for making biochips. A biochip can comprise a nanopore in a membrane (e.g., lipid bilayer) adjacent or in proximity to an electrode. Methods are described for forming the membrane and inserting the nanopore into the membrane. The biochips and methods can be used for nucleic acid (e.g., DNA) sequencing. The present disclosure also describes methods for detecting, sorting, and binning molecules (e.g., proteins) using biochips.
Claims44
A method for forming a lipid bilayer for use in a nanopore sensor, comprising:
- (a) directing an electrically conductive buffer solution in a flow channel comprising an electrode having a lipid material layer dried down thereon, wherein said lipid material layer comprises one or more lipids, and is capable of forming the lipid bilayer;
- (i) directing a bubble through said flow channel and bringing said bubble in contact with said lipid material layer to smooth and/or thin said lipid material layer;
- (ii) flowing a solution comprising a pore protein adjacent to said lipid material layer to deposit said pore protein in said lipid material layer, and thinning said lipid material layer with an additional bubble in said flow channel;
- (b) bringing said buffer solution in contact with said lipid material layer to facilitate the formation of the lipid bilayer on top of said electrode, the pore protein solution and the buffer solution being separated by the additional bubble;
- (c) measuring a current through said electrode to determine if at least a portion of said lipid material layer has formed said lipid bilayer adjacent to said electrode;
- (d) based on the determination of (c), applying a stimulus to said electrode to induce said at least the portion of the material layer to form said lipid bilayer adjacent to said electrode;
- and
- (e) applying a stimulus through said electrode to facilitate the insertion of the pore protein in said lipid bilayer.
5 more elements
A method for forming a lipid bilayer for use in a nanopore sensor, comprising:
- (a) providing a chip comprising a plurality of electrodes and lipid material layers adjacent to said plurality of electrodes, wherein each of said lipid material layers comprises a lipid capable of forming the lipid bilayer;
- (i) directing a bubble through said flow channel and bringing said bubble in contact with said lipid material layer to smooth and/or thin said lipid material layer;
- (ii) flowing a solution comprising a pore protein adjacent to said lipid material layer to deposit said pore protein in said lipid material layer, and thinning said lipid material layer with an additional bubble in said flow channel;
- (b) contacting said lipid material layers with a buffer solution, wherein said buffer solution is electrically conductive, the pore protein solution and the buffer solution being separated by the additional bubble;
- (c) applying a stimulus to at least a subset of said plurality of electrodes to induce said material layers to form lipid bilayers adjacent to the plurality of the electrodes;
- and
- (d) repeating steps (b) and (c), as needed, until at least about 20% of said plurality of electrodes deactivate at a voltage pulse between about −100 millivolts (mV) and −1000 mV applied to said plurality of electrodes;
- and
- (e) applying a stimulus through at least a subset of said plurality of electrodes to facilitate the insertion of said pore protein in each of said lipid bilayers.
6 more elements
Dependencies are inferred from the claim text.