Nanochannel arrays and their preparation and use for high throughput macromolecular analysis

Patent metadata

Field	Value
Publication number	US-10768142-B2
Application number	US-201916367701-A
Country	US
Kind code	B2
Filing date	Mar 28, 2019
Priority date	Jul 25, 2001
Publication date	Sep 8, 2020
Grant date	Sep 8, 2020

Abstract

Nanochannel arrays that enable high-throughput macromolecular analysis are disclosed. Also disclosed are methods of preparing nanochannel arrays and nanofluidic chips. Methods of analyzing macromolecules, such as entire strands of genomic DNA, are also disclosed, as well as systems for carrying out these methods.

First claim

What is claimed is:

1. A method of analyzing at least one macromolecule, comprising the steps of:
transporting at least one macromolecule in a fluid sample into a nanochannel, such that the at least one macromolecule is in an elongated state within the nanochannel;
maintaining the at least one macromolecule in an elongated state within the nanochannel without application of a shear force;
detecting at least one signal transmitted from the at least one elongated macromolecule within the nanochannel; and
correlating the detected signal to at least one property of the at least one macromolecule;
wherein the at least one macromolecule is DNA and the detected signal is correlated to a base pair sequence of said DNA.

2. The method of claim 1 , wherein the at least one macromolecule is transported into the nanochannel by applying an electric current.

3. The method of claim 1 , wherein the trench width of the nanochannel is less than about 150 nanometers.

4. The method of claim 1 , wherein the trench depth of the nanochannel is less than about 200 nanometers.

5. The method of claim 1 , wherein the width of the nanochannel is substantially constant along the length of the nanochannel.

6. The method of claim 1 , wherein the nanochannel is in the material of a surface.

7. The method of claim 1 , wherein the nanochannel is surmounted by sealing material to render the nanochannel at least substantially enclosed.

8. The method of claim 7 , wherein the sealing material is optically transparent.

9. The method of claim 8 , wherein at least one signal is transmitted through the optically transparent sealing material.

10. The method of claim 1 , wherein the at least one detected signal is correlated to at least one of the following properties: length, conformation, and chemical composition.

11. The method of claim 1 , wherein the at least one macromolecule has an elongated length in nanochannel of greater than 150 nanometers.

12. The method of claim 1 , wherein the at least one macromolecule is DNA having greater than 10 base pairs.

13. The method of claim 1 , further comprising the step of illuminating the at least one macromolecule within the nanochannel.

14. A method of analyzing at least one macromolecule, comprising the steps of:
transporting at least one macromolecule in a fluid sample into a nanochannel, such that the at least one macromolecule is in an elongated state within the nanochannel;
maintaining the at least one macromolecule in an elongated state within the nanochannel without application of a shear force;
detecting at least one signal transmitted from the at least one elongated macromolecule within the nanochannel; and
correlating the detected signal to at least one property of the at least one macromolecule;
wherein the at least one macromolecule is a restriction fragment length polymorphism.

15. A method of analyzing at least one macromolecule, comprising the steps of:
transporting at least one macromolecule in a fluid sample into a nanochannel, such that the at least one macromolecule is in an elongated state within the nanochannel;
maintaining the at least one macromolecule in an elongated state within the nanochannel without application of a shear force;
detecting at least one signal transmitted from the at least one elongated macromolecule within the nanochannel; and
correlating the detected signal to at least one property of the at least one macromolecule;
wherein the at least one macromolecule is at least one chromosome.

16. A method of analyzing at least one macromolecule, comprising the steps of:
transporting at least one macromolecule in a fluid sample into a nanochannel, such that the at least one macromolecule is in an elongated state within the nanochannel;
maintaining the at least one macromolecule in an elongated state within the nanochannel without application of a shear force;
detecting at least one signal transmitted from the at least one elongated macromolecule within the nanochannel; and
correlating the detected signal to at least one property of the at least one macromolecule;
wherein the at least one chromosome is analyzed to determine the presence of at least one single nucleotide polymorphism.

Assignees

• Univ Princeton

Inventors

• Austin Robert H
• Yu Zhaoning
• Tegenfeldt Jonas O
• Chou Stephen Y
• Cao Han

Classifications

• B01L2300/0627

  Sensor or part of a sensor is integrated · CPC title

• G01N33/48721

  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

• B01L3/502707

  characterised by the manufacture of the container or its components · CPC title

• G01N21/6428

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title

• Y10S977/704

  Nucleic acids, e.g. DNA or RNA · CPC title