Nanopore sequencing using n-mers
The invention relates to devices and methods for nanopore sequencing. The invention provides devices having an array of nanopores. The devices comprise an upper fluidic region in fluidic contact with the nanopores. The upper fluidic region comprises and upper electrode. Below each nanopore, and in fluidic contact with each nanopore is a discrete fluidic region. There is no direct fluidic contact between these discrete fluidic regions. The devices have an advantage of lower levels of electrical cross-talk between nanopores in the array for improved nucleic acid sequencing.
1. A device for nucleic acid sequencing comprising:
an array of at least 100 nanopores,
above the array of nanopores, in fluidic contact with the array of nanopores, an upper fluidic region comprising an upper electrode, the upper fluidic region comprising nucleic acid molecules,
below the array of nanopores, an array of discrete fluidic regions, each discrete fluidic region in fluidic contact with a nanopore, and each discrete fluidic region comprising a lower electrode, wherein there is no direct fluidic contact between the discrete fluidic regions, and
below the array of discrete fluidic regions, an array of electronic circuits, each electronic circuit comprising an amplifier in electrical contact with the fluidic region above it,
wherein voltage is applied between the upper electrode and the lower electrodes, resulting in translocation of a nucleic acid molecule through a plurality of the nanopores, and whereby a measured electrical signal is used to determine sequences of a plurality of nucleic acid molecules.
2. The device of claim 1 wherein the device comprises a semiconductor substrate in which the electronic circuits are formed.
3. The device of claim 2 wherein the electronic circuits comprise CMOS circuits.
4. The device of claim 1 wherein the electronic circuits further comprise analog to digital converters, memory, or clock circuits.
5. The device of claim 1 wherein each discrete reservoir has two electrodes, one acting as a drive electrode, and one acting as a measurement electrode.
6. The device of claim 1 wherein the nanopores comprise protein nanopores.
7. The device of claim 6 wherein the protein nanopores comprise alpha-hemolysin proteins.
8. The device of claim 6 wherein the protein nanopores comprise MspA proteins.
9. The device of claim 1 wherein the protein nanopores are within biological membranes.
10. The device of claim 1 wherein the nanopores comprise solid state nanopores.
11. The device of claim 1 wherein the nanopores comprise hybrid solid state-protein nanopores.
12. The device of claim 1 wherein each discrete fluidic region has a cross sectional dimension from 1 micron to 10 microns.
13. The device of claim 1 wherein the upper fluidic region further comprises enzymes for control of translocation rate.
14. The device of claim 13 wherein the enzymes for control of translocation rate comprise polymerases.
15. The device of claim 13 wherein the enzymes for control of translocation rate comprise exonucleases.
16. The device of claim 13 wherein the enzymes for control of translocation rate comprise helicases.
17. The device of claim 1 wherein the nucleic acid comprises DNA.
18. The device of claim 1 wherein the device comprises a substrate comprising a semiconductor component bound to an insulator component.
19. The device of claim 1 comprising from 100 to 10,000 nanopores.