Nanopump device for analyzing DNA sequences

• US 20040038260A1
• Filed: 04/17/2003
• Published: 02/26/2004
• Est. Priority Date: 04/18/2002
• Status: Abandoned Application

• Current Assignee: Imedd, Inc.
• Original Assignee: Imedd, Inc.
• Inventors: Martin, Francis J., Walczak, Robbie J.
• Application Number:
  US10/417,859
• Publication Number:
  US 20040038260A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2565/125 Electrophoretic separation
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: C12Q 2565/631 being a biochannel or pore

Abstract

The invention includes a device for detecting a target nucleic acid sequence contained in a solution of solution of nucleic acid fragments. The device includes a chamber, and a membrane disposed in said chamber and having a channel extending between an upstream chamber region, where the said channel has a selected minimum cross-sectional dimension in the range between 2 and 100 nm. Attached to a wall portion of the channel, is a capture nucleic acid having a sequence complementary to the target sequence. Upstream and downstream electrodes disposed in the upstream and downstream chamber regions, respectively, are in contact with electrolyte solution placed in the corresponding chamber regions. A controller in the device includes a power source operatively connected to the electrodes for applying a selected voltage potential across the channel, to move individual nucleic acid sequences contained in the solution through the channel, where the sequences can hybridize to complementary target sequences bound to the channel wall portion.

Claims

1. A device for detecting a target nucleic acid sequence contained in a solution of solution of nucleic acid fragments and having a selected pH, comprising:

a chamber,

a membrane disposed in said chamber and having a channel extending between an upstream chamber region adapted to hold the electrolyte solution of such different-length fragments, and a downstream chamber region adapted to hold an electrolyte solution, where said channel has a selected minimum cross-sectional dimension in the range between 2 and 100 nm and a net surface charge within a given pH range that includes the selected solution pH,

attached to a wall portion of the channel, a capture nucleic acid having a sequence complementary to the target sequence,

upstream and downstream electrodes disposed in said upstream and downstream chamber regions, respectively, for contacting solution placed in the corresponding chamber regions,

a controller including a power source operatively connected to said electrodes for applying a selected voltage potential across said channel, to move individual nucleic acid sequences contained in the solution through said channel, where said sequences can hybridize to complementary target sequences bound to the channel wall portion.

2. The device of claim 1, wherein said controller further includes a voltage regulator for regulating the voltage applied across said channel, to effect selective release of the target sequence from the capture nucleic acid, based on the degree of complementarity between the target sequences and the capture nucleic acid.

3. The device of claim 1, wherein said membrane includes an array of channels, each having attached to a wall portion thereof, a capture nucleic acid with a selected sequence complementary to a selected one of a plurality of different sequences.

4. The device of claim 3, wherein one of said electrodes includes a plurality of electrode elements, each associated with one of said membrane channels, and said controller is operatively connected to each of said electrode elements to apply and regulate the voltage applied across each of the channels.

5. The device of claim 1, wherein said controller is effective to place across the electrodes, a voltage potential effective to move nucleic acid fragments electrophoretically through said channel.

6. The device of claim 1, wherein said channel has a minimum dimension in the 2-25 nm range.

7. A method for detecting a target nucleic acid sequence contained in an electrolyte solution of nucleic acid fragments and having a selected pH, comprising

placing the solution in a chamber having a membrane disposed therein, said membrane having a channel extending between an upstream chamber region adapted to hold the electrolyte solution of such different-length fragments, and a downstream chamber region adapted to hold an electrolyte solution, where said channel (i) has a selected minimum cross-sectional dimension in the range between 2 and 100 nm, (ii) a net surface charge within a given pH range that includes the selected solution pH, and (iii), attached to a wall portion of the channel, a capture nucleic acid having a sequence complementary to the target sequence,

applying across the channel, a voltage potential sufficient to move nucleic acid sequences in said solution through said channel, where said sequences can hybridize to complementary target sequences bound to the channel wall portion,

releasing captured target sequence from said channel by applying a voltage potential across said channel effect to dissociate hybridized nucleic acids, and

detecting released target sequences.

8. The method of claim 7, wherein said releasing includes applying a voltage across said channel effective to selectively release target sequences from the capture nucleic acid, based on the degree of complementarity between the target sequence and the capture nucleic acid.

9. The method of claim 7, wherein said membrane includes an array of channels, each having attached to a wall portion thereof, a capture nucleic acid with a selected sequence complementary to a selected one of a plurality of different sequences.

10. The method of claim 9, wherein one of said electrodes includes a plurality of electrode elements, each associated with one of said membrane channels, said controller is operatively connected to each of said electrode elements to apply and regulate the voltage applied across each of the channels, and said releasing includes applying a voltage across each of said channels, individually, a voltage potential effective to selectively release target sequences from the capture nucleic acid, based on the degree of complementarity between the target sequence and the capture nucleic acid.