Enrichment of nucleic acid targets

Patent metadata

Field	Value
Publication number	US-11795497-B2
Application number	US-202117227121-A
Country	US
Kind code	B2
Filing date	Apr 9, 2021
Priority date	Feb 2, 2004
Publication date	Oct 24, 2023
Grant date	Oct 24, 2023

Abstract

Methods and apparatus providing for the isolation of an unknown mutation from a sample comprising wild type nucleic acids and mutated nucleic acids through the application of time-varying driving fields and periodically varying mobility-altering fields to the sample within in an affinity matrix.

First claim

What is claimed is:

1. A method of sequencing a target molecule comprising:
(a) isolating the target molecule from a sample, wherein the isolating comprises
(i) applying a time-varying driving field to the sample in the presence of an affinity matrix comprising immobilized probes, the probes having a binding affinity for the target molecule;
(ii) applying a periodically varying, mobility-altering field that modifies the binding affinity;

wherein application of the time-varying driving field and the periodically varying, mobility-altering field isolates the target molecule from the sample; and
(b) subjecting the target molecule to sequencing.

2. The method of claim 1 , wherein the periodically varying mobility-altering field is a temperature gradient.

3. The method of claim 1 , wherein the time-varying driving field comprises an electric field.

4. The method of claim 1 , wherein the time-varying driving field varies direction with time.

5. The method of claim 1 , wherein the immobilized probes have a binding affinity for the target molecule that is greater than for any non-target molecule in the sample.

6. The method of claim 1 , wherein the target molecule is a nucleic acid and wherein the immobilized probes each comprise a nucleic acid that is complementary to at least a portion of the target molecule.

7. The method of claim 1 , wherein the sequencing is nucleic acid sequencing.

8. The method of claim 7 , wherein the nucleic acid sequencing is Sanger sequencing, sequencing by synthesis, pyrosequencing, nanopore sequencing, single-molecule sequencing, or fluorescence-based sequencing.

9. The method of claim 1 , wherein the sample is a biological sample.

10. The method of claim 9 , wherein the biological sample is a tissue, blood, sputum, sweat, urine, feces, tears, or aspirate sample.

11. The method of claim 1 , wherein the isolating is performed using a device comprising the affinity matrix.

12. The method of claim 1 , wherein the isolating is performed using a column or capillary comprising the affinity matrix.

13. The method of claim 1 , wherein the sequencing is performed using a sequencing device.

14. A method of sequencing a target molecule comprising:
(a) isolating a target molecule from a sample using scodaphoresis; and
(b) subjecting the target molecule to sequencing.

15. The method of claim 14 , wherein scodaphoresis comprises application of a time-varying driving field and a periodically varying, mobility-altering field.

16. The method of claim 14 , wherein scodaphoresis comprises an affinity matrix comprising immobilized probes, wherein the probes have a binding affinity for the target molecule.

Assignees

• Quantum Si Inc

Inventors

• Marziali Andrea
• Despotovic Milenko
• Pel Joel

Classifications

• C12Q1/6832 Primary

  Enhancement of hybridisation reaction · CPC title

• B01D29/05

  supported · CPC title

• B01D35/027

  rigidly mounted in or on tanks or reservoirs (B01D35/04 takes precedence) · CPC title

• B01D35/0273

  Filtering elements with a horizontal or inclined rotation or symmetry axis submerged in tanks or reservoirs · CPC title

• B01L3/502753

  characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation · CPC title