Massively parallel DNA sequencing apparatus

Patent metadata

Field	Value
Publication number	US-10712334-B2
Application number	US-201716073706-A
Country	US
Kind code	B2
Filing date	Jan 27, 2017
Priority date	Jan 28, 2016
Publication date	Jul 14, 2020
Grant date	Jul 14, 2020

Abstract

A DNA or genome sequencing structure is disclosed. The structure includes an electrode pair, each electrode having a tip-shaped end, the electrodes separated by a nanogap defined by facing tip-shaped ends; at least one conductive island deposited at or near each tip-shaped end; and a biomolecule having two ends, each end attached to the conductive islands in the electrode pair such that one biomolecule bridges over the nanogap in the electrode pair, wherein nucleotide interactions with the biomolecule provides electronic monitoring of DNA or genome sequencing without the use of a fluorescing element.

First claim

We claim:

1. A DNA or genome sequencing structure comprising:
(a) an electrode pair comprising a first metal, each electrode tapering to a tip-shaped end, with the electrodes separated by a nanogap defined by the tip-shaped ends facing one another;
(b) at least one conductive island deposit of a second metal on each electrode located at or near each tip-shaped end of each electrode; and
(c) a biomolecule having two ends, each end attached to the at least one conductive island on each electrode such that one biomolecule bridges the nanogap,
wherein the first and second metals are different, and
wherein the biomolecule completes a circuit configured for electronic monitoring of DNA or genome sequencing without the use of a fluorescing element.

2. The structure of claim 1 , wherein the first metal is platinum (Pt), palladium (Pd), rhodium (Rh), or titanium (Ti).

3. The structure of claim 1 , wherein the second metal is gold (Au).

4. The structure of claim 3 , wherein the at least one conductive island deposit comprises a gold (Au) electrodeposit.

5. The structure of claim 3 , wherein the at least one conductive island deposit comprises a gold (Au) nanoparticle.

6. The structure of claim 3 , wherein the at least one conductive island comprises a cylindrically shaped nano-pillar.

7. The structure of claim 6 , wherein the nano-pillar measures less than 20 nm in diameter and less than 25 nm in height.

8. The structure of claim 6 , wherein the nano-pillar measures less than 7 nm in diameter and less than 10 nm in height.

9. The structure of claim 3 , wherein each end of the biomolecule is attached to the at least one conductive island through thiol-gold (Au) binding or gold binding proteins.

10. The structure of claim 1 , wherein the at least one conductive island deposit comprises a nano-tip shaped or nano-pillar shaped conductive island having an exposed dimension of less than 20 nm.

11. The structure of claim 10 , wherein the nano-tip or nano-pillar comprises a branched or porous surface, having a porosity of at least 30% so as to increase the surface area of an exposed surface of the nano-tip or nano-pillar by at least by 10% as compared to the nano-tip or nano-pillar structure after electrodeposition or electroless deposition on each electrode.

12. The structure of claim 1 , wherein the structure comprises a plurality of layers of electrode pairs so as to form a three-dimensional array.

13. The structure of claim 12 , wherein the electrode pairs are connected to one another such that one electrode from each pair are ganged together by a common lead wire and each of the other electrodes in each pair are left unconnected to one another, enabling independent and sequential interrogation of each electrode pair.

14. The structure of claim 1 , wherein each end of the biomolecule is attached to the at least one conductive island through antibody-antigen coupling or streptavidin-biotin coupling.

15. A genome or DNA sequencing system comprising:
(a) the DNA or genome sequencing structure of claim 1 ; and
(b) a chamber encasing the structure and defining a microfluidic subsystem usable to supply biomolecules, nucleotides, PBS, or water solutions to the electrode pair.

16. A method of making a genome or DNA sequencing device, said method comprising the steps of:
(a) disposing an array of electrode pairs on a substrate, each electrode within a given pair of electrodes having a tip-shaped end, with the electrodes in each pair separated by a nanogap defined by tip-shaped ends facing one another;
(b) electrodepositing gold (Au) at each tip-shaped end by applying a voltage to the electrode pair whereby high current density in the region of each tip-shaped end of each electrode directs preferential electrodeposition of gold (Au) to each tip-shaped end to form a gold (Au) nano-tip on each electrode; and
(c) attaching each end of a biomolecule having two ends to the gold (Au) nano-tips such that one biomolecule bridges over each nanogap in each electrode pair.

17. The method of claim 16 , wherein the electrode pairs comprise platinum (Pt), palladium (Pd) or rhodium (Rh).

18. The method of claim 16 , wherein the substrate comprises silicon (Si) with a SiO 2 insulator surface.

19. The method of claim 16 , further comprising heat treating the array of electrode pairs after step (b) at from about 200 to about 800° C. to induce additional diffusional bonding of gold (Au) to the metal electrodes.

20. The method of claim 16 , further comprising patterning a passivation layer over the array of electrode pairs, prior to step (c), in order to cover undesired errant Au deposits in locations other than the tip-shaped ends of the electrodes.

21. A method of making a genome or DNA sequencing device, said method comprising the steps of:
(a) disposing an array of electrode pairs on a substrate, each electrode within a given pair of electrodes having an end, with the electrodes in each pair separated by a nanogap defined by the ends of the electrodes facing one another;
(b) disposing a mask resist layer over the electrodes;
(c) nano-patterning the mask resist layer to form openings, one opening per electrode, wherein each opening is at or near each nanogap;
(d) electrodepositing gold (Au) on each electrode through each opening to form gold (Au) nano-pillars; and
(e) attaching each end of a biomolecule having two ends to the gold (Au) nano-pillars such that one biomolecule bridges over each nanogap in each electrode pair.

22. The method of claim 21 , wherein the electrode pairs comprise platinum (Pt), palladium (Pd) or gold (Au).

23. The method of claim 21 , wherein the substrate comprises silicon (Si) with a SiO 2 insulator surface.

24. The method of claim 21 , wherein the resist layer comprises PMMA or hydrogen silsesquioxane (HSQ).

25. The method of claim 21 , wherein the gold (Au) nano-pillars grow within each opening with each opening filled by the electrodeposition of gold (Au).

Assignees

• Roswell Biotechnologies Inc

Inventors

• Choi Chulmin
• Jin Sungho
• Mola Paul W
• Merriman Barry L

Classifications

• C12Q2565/607

  being a sensor, e.g. electrode · CPC title

• G01N27/3278 Primary

  involving nanosized elements, e.g. nanogaps or nanoparticles (nanopores G01N33/48721; magnetic beads G01N27/745) · CPC title

• C12Q1/6869

  Methods for sequencing · CPC title

• G01N33/48721 Primary

  Investigating individual macromolecules, e.g. by translocation through nanopores (Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68) · CPC title

• Y02E60/50

  Fuel cells · CPC title