Molecule detection system on a solid support

• US 9,222,134 B2
• Filed: 06/17/2013
• Issued: 12/29/2015
• Est. Priority Date: 09/17/2010
• Status: Active Grant

• Current Assignee: Illumina Incorporated
• Original Assignee: Illumina Incorporated
• Inventors: Mann, Tobias
• Primary Examiner(s): Lin, Jerry
• Application Number:
  US13/919,910
• Publication Number:
  US 20130281321A1
• Time in Patent Office: 925 Days
• Field of Search: 702/19
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: G01N 33/50 Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing measuring or testing processes involving enzymes or microorganisms, compositions or test papers therefor; processes for forming such compositions, condition responsive control in microbiological or enzymological processes C12Q
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G16B 25/00 ICT specially adapted for hybridisation; ICT specially adapted for gene or protein expression
• CPC Class Codes: G16C 20/20 Identification of molecular entities, parts thereof or of chemical compositions

Abstract

Disclosed herein are compositions and methods used for detecting different types of molecules associated with a site on a solid support.

Claims

1. A system of detecting molecules, said system comprising:

(a) a solid support comprising molecules associated with a site on the solid support, wherein the site comprises at least two different types of molecules comprising nucleic acids;

(b) a detector configured to detect at least one signal corresponding to aggregates of molecules comprising nucleic acids at the site;

(c) a first data processing module configured to provide an initial estimate of the amount of signal corresponding to each of the at least two different types of molecules comprising nucleic acids at the site;

(d) a second data processing module configured to calculate the amount of signal corresponding to each of the at least two different types of molecules comprising nucleic acids at the site using the initial estimate, thereby obtaining a signal estimate; and

(e) a third data processing module configured to iteratively update the initial estimate until the signal estimates converge, thereby detecting molecules associated with the site.

2. The system of claim 1, wherein the solid support comprises glass, plastic, carbon, metal, a resin or silicon.

3. The system of claim 2, wherein the plastic is selected from the group consisting of acrylics, polystyrenes, copolymers of styrene, polypropylene, polyethylene, polybutylene, polyurethanes, and Teflon.

4. The system of claim 1, wherein the solid support is a planar solid support.

5. The system of claim 1, wherein the solid support comprises a flow-cell.

6. The system of claim 1, wherein the solid support comprises wells.

7. The system of claim 1, wherein the solid support comprises beads attached to the at least two different types of molecules comprising nucleic acids.

8. The system of claim 1, further comprising a robotic automation system.

9. The system of claim 8, wherein the robotic automation system comprises a device selected from the group consisting of a robotic arm, a plate handler, an automated lid handler, a fluid handling device, a well loading block, a heating/cooling system, and a reagent rack.

10. The system of claim 1, wherein the detector comprises an energy source configured to provide excitation energy to one or more sites on the solid support.

11. The system of claim 1, wherein the detector further comprises a CCD camera, or CMOS.

12. The system of claim 1, wherein the first data processing module is configured to perform a principal component analysis (PCA) to determine the initial estimate of the amount of signal corresponding to each of the at least two different types of molecules comprising nucleic acids at the site.

13. The system of claim 1, wherein the third data processing module is configured to perform a numerical optimization algorithm.

14. The system of claim 13, wherein the numerical optimization algorithm is based on an iterative map search or Fienup'"'"'s iteration map.

15. The system of claim 1, wherein a data processing module is configured to obtain sequence data for one or more of the at least two different molecules comprising nucleic acids.

16. The system of claim 15, wherein sequence data is obtained by a sequencing-by-synthesis process.

17. The system of claim 1, wherein the molecule detection system is configured to identify the nucleotide sequence of a target region of a nucleic acid.

18. The system of claim 1, wherein the nucleic acids comprise a first subpopulation of nucleic acids having first target regions and a second subpopulation of nucleic acids having second target regions, wherein the first target regions are identical to one another and the second target regions are identical to one another, and the second target regions are a variant of the first target regions.

19. The system of claim 18, wherein the nucleotide sequence of said target region of the nucleic acids of the first subpopulation has at least 3 nucleotides that are different as compared to the nucleotide sequence of said variant of the target region of the nucleic acids of the second subpopulation.

20. The system of claim 1, wherein the nucleic acids comprise alleles of a genetic locus.

21. A system of detecting molecules, said system comprising:

(a) a solid support comprising molecules associated with a site on the solid support, wherein the site comprises at least two different types of molecules, wherein the at least two different types of molecules comprise nucleic acids;

(b) a detector configured to detect at least one signal corresponding to aggregates of molecules comprising nucleic acids at the site;

(c) a first data processing module configured to provide an initial estimate of the fraction of signal corresponding to each of the at least two different types of molecules comprising nucleic acids at the site;

(d) a second data processing module configured to calculate the fraction of signal corresponding to each of the at least two different types of molecules comprising nucleic acids at the site using the initial estimate, thereby obtaining a fraction estimate; and

(e) a third data processing module configured to iteratively update the initial estimate until the fraction estimates converge, thereby detecting molecules associated with the site.

22. A system for detecting molecules, comprising:

(a) a solid support comprising molecules associated with a site on the solid support such that the molecules are detected in aggregate during a detection step, wherein the site comprises at least two different types of molecules, wherein the molecules comprise nucleic acids;

(b) means for detecting a signal corresponding to the aggregate of molecules comprising nucleic acids at the site;

(c) means for providing an initial estimate of the amount of signal corresponding to each of the at least two different types of molecules comprising nucleic acids at the site;

(d) means for calculating the amount of signal corresponding to each of the at least two different types of molecules comprising nucleic acids at the site using the initial estimate, thereby obtaining a signal estimate; and

(e) means for iteratively updating the initial estimate until the signal estimates converge, thereby detecting molecules associated with the site.