Compositions, devices, systems, and methods for using a nanopore
Devices and methods that can detect and control an individual polymer in a mixture is acted upon by another compound, for example, an enzyme, in a nanopore are provided. The devices and methods also determine (˜>50 Hz) the nucleotide base sequence of a polynucleotide under feedback control or using signals generated by the interactions between the polynucleotide and the nanopore. The invention is of particular use in the fields of molecular biology, structural biology, cell biology, molecular switches, molecular circuits, and molecular computational devices, and the manufacture thereof.
1. A method for analyzing labeled polymers, the method being performed using a system comprising:
a plurality of independently addressable elements, each independently addressable element comprising a thin film separating a cis compartment from a trans compartment, wherein the thin film of each independently addressable element comprises a pore therein, wherein the cis compartment is common to the plurality of independently addressable elements and comprises a common cis electrode, wherein the trans compartment of each independently addressable element is separate from trans compartments of other independently addressable elements and comprises a trans electrode; and
a power source electrically coupled to the common cis electrode and the trans electrodes, wherein the power source, the common cis electrode and the trans electrodes are configured to apply a potential difference across the thin films of the plurality of independently addressable elements,
the method comprising, for each independently addressable element:
(a) providing a labeled polymer to a solution in the cis compartment;
(b) applying a potential difference across the thin film to translocate labels of the labeled polymer through the pore to a solution in the trans compartment;
(c) measuring a transport property during translocation of the labels from the cis compartment to the trans compartment; and
(d) determining a sequence of the labeled polymer based on the transport property measured solely by the independently addressable element, wherein the sequence comprises two or more different types of monomers of the labeled polymer.
2. The method according to claim 1 , wherein the polymer has been extended by a polymerase.
3. The method according to claim 1 , wherein the labeled polymer comprises nucleobases.
4. The method according to claim 1 , wherein the labeled polymer comprises a phosphate backbone.
5. The method according to claim 1 , wherein the labeled polymer comprises a duplex region, and wherein applying a potential difference across the pore unzips the duplex region.
6. The method according to claim 1 , wherein the transport property comprises impedance, current, resistance, or a combination thereof.
7. The method according to claim 1 , wherein the pore of each independently addressable element is a nanopore.
8. The method according to claim 1 , wherein the pore of each independently addressable element is a solid state pore.
9. The method according to claim 1 , wherein the pore of each independently addressable element is a biological pore.
10. The method according to claim 1 , wherein the thin film is an inorganic membrane, a solid state membrane, or a molecular bilayer.
11. A system for analyzing labeled polymers, the system comprising:
a plurality of independently addressable elements, each independently addressable element comprising a thin film separating a cis compartment from a trans compartment, wherein the thin film of each independently addressable element comprises a pore therein, wherein the cis compartment is common to the plurality of independently addressable elements and comprises a common cis electrode, wherein the trans compartment of each independently addressable element is separate from trans compartments of other independently addressable elements and comprises a trans electrode;
a power source electrically coupled to the common cis electrode and the trans electrodes, wherein the power source, the common cis electrode and the trans electrodes are configured to apply a potential difference across the thin films of the plurality of independently addressable elements; and
a processor coupled to the power source and programmed to, for each independently addressable element:
(a) control the power source to apply a potential difference across the thin film to translocate labels of a labeled polymer present in a solution in the cis compartment through the pore to a solution in the trans compartment;
(b) measure a transport property during translocation of the labels from the cis compartment to the trans compartment; and
(c) determine a sequence of the labeled polymer based on the transport property measured solely by the independently addressable element, wherein the sequence comprises two or more different types of monomers of the labeled polymer.
12. The system of claim 11 , wherein the labeled polymer comprises nucleobases.
13. The system of claim 11 , wherein the transport property comprises impedance, current, resistance, or a combination thereof.
14. The system of claim 11 , wherein the pore of each independently addressable element is a nanopore.
15. The system of claim 14 , wherein the nanopore has an aperture size of from 1 to 4 nm.
16. The system of claim 11 , wherein the pore of each independently addressable element is a solid state pore.
17. The system of claim 11 , wherein the pore of each independently addressable element is a biological pore.
18. The system of claim 11 , wherein the thin film is an inorganic membrane, a solid state membrane, or a molecular bilayer.