IP Library Granted Patent US 8,906,611
Granted Patent B2
US 8,906,611 · App. 13/190,947 · Granted Dec 9, 2014

Devices and methods for immobilizing nucleic acids

Inventor: Wenlong Jiang (Madison, WI)
Assignee: OpGen, Inc.
B01L3/502761C12Q2523/31B01L2400/0427C12Q1/689B01L2200/0663C12Q1/6825
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Quick Facts
Patent No.
US 8,906,611
App. No.
13/190,947
Granted
Dec 9, 2014
Kind
B2
Abstract

The present invention generally relates to devices and methods for immobilizing nucleic acids on a substrate. In certain embodiments, devices of the invention include a voltage source, and a substrate coupled to the voltage source, in which hydrophobicity of the substrate changes in response to an applied electric field and a surface of the substrate is coated with a substance that retains nucleic acids.

Claims (24)

1. A method for elongating a nucleic acid, the method comprising:

providing a device comprising a voltage source coupled to a controller, a fluid reservoir, and a charged substrate operably associated with the voltage source and the fluid reservoir, wherein a hydrophobicity of the charged substrate changes in response to an applied electric field and the substrate is coated with a substance that retains the nucleic acid;

temporarily applying an electric field via the controller to temporarily decrease the hydrophobicity of the charged substrate, thereby causing a sample fluid comprising the nucleic acid in the fluid reservoir to temporarily flow from the reservoir onto the charged substrate;

maintaining the electric field via the controller so that the sample fluid remains on the charged substrate for a time sufficient to allow a sufficient amount of the nucleic acid to interact with the charged substrate and become elongated and fixed on the substrate; and

turning-off the electric field via the controller to thereby increase the hydrophobicity of the charged substrate and cause the sample fluid to flow back into the reservoir while the elongated and fixed nucleic acid remains on the charged substrate.

2. The method according to claim 1 , wherein the nucleic acid is DNA.

3. The method according to claim 1 , wherein the nucleic acid is from a microorganism.

4. The method according to claim 3 , wherein the microorganism is a bacterium.

5. A method for characterizing a nucleic acid, the method comprising:

providing a device comprising a voltage source coupled to a controller, one or more fluid reservoirs, and a charged substrate operably associated with the voltage source and the one or more reservoirs, wherein a hydrophobicity of the substrate changes in response to an applied electric field and the charged substrate is coated with a substance that retains nucleic acids;

temporarily applying an electric field via the controller to temporarily decrease the hydrophobicity of the charged substrate, thereby causing a sample fluid comprising the nucleic acid to temporarily flow from the fluid reservoir onto the charged substrate, wherein the fluid reservoir is fluidically coupled to the device;

maintaining the electric field via the controller so that the sample fluid remains on the charged substrate for a time sufficient to allow a sufficient amount of the nucleic acid to interact with the charged substrate and become elongated and fixed on the substrate so that the nucleic acid remains accessible for enzymatic reactions;

turning-off the electric field via the controller to thereby increase the hydrophobicity of the charged substrate and cause the sample fluid to flow back into the reservoir while the elongated and fixed nucleic acid remains on the charged substrate;

intermittently re-applying the electric field via the controller to thereby intermittently decrease the hydrophobicity of the charged substrate to cause reagents to flow to and from the substrate to wash, enzymatically digest, and stain the nucleic acid to obtain one or more restriction digests of the nucleic acid, wherein the reagents are in individual fluid reservoirs, the individual fluid reservoirs are fluidically coupled to the device, and turning-off the electric field via the controller increases the hydrophobicity of the charged substrate and causes the reagents to flow back into the respective individual reservoir; and

imaging the restriction digests, thereby characterizing the nucleic acid.

6. The method according to claim 5 , further comprising constructing an optical map from the restriction digests.

7. The method according to claim 5 , wherein the nucleic acid is DNA.

8. The method according to claim 5 , wherein the nucleic acid is from a microorganism.

9. The method according to claim 8 , wherein the microorganism is a bacterium.

10. The method according to claim 9 , wherein the nucleic acid comprises substantially all genomic DNA of the bacterium.

11. The method according to claim 9 , wherein the nucleic acid comprises a transcriptome of the bacterium.

12. The method according to claim 9 , wherein the bacterium is at least one species selected from the group consisting of Escherichia coli ( E. coli ) and Staphylococcus aureus ( S. aureus ).

13. The method according to claim 12 , wherein the S. aureus is a community-acquired methicillin-resistant strain of S. aureus.

14. The method according to claim 12 , wherein the S. aureus is a hospital-acquired methicillin-resistant strain of S. aureus.

Assignments (3)
RELEASE OF SECURITY INTEREST Recorded Feb 3, 2021
From: MERCK GLOBAL HEALTH INNOVATION FUND, LLC
To: OPGEN, INC.; ADVANDX, INC.
Reel/Frame 055209/0242 →
SECURITY INTEREST Recorded Aug 17, 2015
From: OPGEN, INC.; ADVANDX, INC.
To: MERCK GLOBAL HEALTH INNOVATION FUND, LLC
Reel/Frame 036377/0129 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 19, 2011
From: JIANG, WENLONG
To: OPGEN, INC.
Reel/Frame 026928/0281 →
Continuity (1)
Related Publication 20130029347A1 · Jan 31, 2013