Patterned flow-cells useful for nucleic acid analysis

• US 8,778,848 B2
• Filed: 06/08/2012
• Issued: 07/15/2014
• Est. Priority Date: 06/09/2011
• Status: Active Grant

• Current Assignee: Illumina Incorporated
• Original Assignee: Illumina Incorporated
• Inventors: Lin, Shengrong, Wu, Yir-Shyuan, Gunderson, Kevin, Moon, John A.
• Primary Examiner(s): Vivlemore, Tracy
• Assistant Examiner(s): Weiler, Karen S
• Application Number:
  US13/492,661
• Publication Number:
  US 20120316086A1
• Time in Patent Office: 767 Days
• Field of Search: None
• US Class Current: 506/23
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: G01N 27/447 using electrophoresis

Abstract

Provided is a surface having metal regions and an interstitial region having a composition that differs from the metal regions, wherein a continuous gel layer coats the surface across the metal regions and the interstitial regions. Nucleic acids or other analytes can be attached to the continuous gel layer such that a greater amount is attached over the metal regions than over the interstitial region. Also provided are methods for making such surfaces. Methods are also provided for making an array of nucleic acids or other analytes using such surfaces.

Claims

1. A method of making an analyte array, comprising

(a) providing a surface comprising mask regions and transparent regions, the mask regions having a composition that blocks transmittance of radiation in a first part of the electromagnetic spectrum and transparent regions having a composition that transmits radiation in the first part of the electromagnetic spectrum, wherein a continuous gel layer coats the surface across the mask regions and the transparent regions;

(b) contacting the continuous gel layer with a fluid comprising a population of different analytes under conditions wherein analytes from the population become attached to the gel layer and wherein a first subpopulation of the analytes attach to portions of the gel layer that coat the mask regions and a second subpopulation of the analytes attach to portions of the gel layer that coat the transparent regions; and

(c) irradiating the surface with radiation in the first part of the electromagnetic spectrum, thereby selectively modifying the analytes of one of the subpopulations compared to the analytes of the other subpopulation,

wherein the selectively modifying comprises selectively removing the analytes of the second subpopulation compared to the analytes of the first subpopulation.

2. The method of claim 1, further comprising removing the mask regions from the surface under conditions wherein the analytes remain attached to the gel layer on the surface.

3. The method of claim 1, wherein the removing comprises photo-cleaving the analytes of the second population using radiation in the first part of the electromagnetic radiation spectrum.

4. The method of claim 3, wherein the analytes of the first subpopulation are masked from the photo-cleaving by the mask regions.

11. The method of claim 1, wherein the transparent regions comprise glass, plastic or silica.

5. A method of making an analyte array, comprising

(a) providing a surface comprising mask regions and transparent regions, the mask regions having a composition that blocks transmittance of radiation in a first part of the electromagnetic spectrum and transparent regions having a composition that transmits radiation in the first part of the electromagnetic spectrum, wherein a continuous gel layer coats the surface across the mask regions and the transparent regions;

(b) contacting the continuous gel layer with a fluid comprising a population of different analytes under conditions wherein analytes from the population become attached to the gel layer and wherein a first subpopulation of the analytes attach to portions of the gel layer that coat the mask regions and a second subpopulation of the analytes attach to portions of the gel layer that coat the transparent regions; and

(c) irradiating the surface with radiation in the first part of the electromagnetic spectrum, thereby selectively modifying the analytes of one of the subpopulations compared to the analytes of the other subpopulation,

wherein the analytes comprise nucleic acids,

further comprising amplifying the nucleic acids on the portions of the gel layer that coat the mask regions.

6. The method of claim 5, further comprising removing the mask regions from the surface under conditions wherein the products of the amplifying of the nucleic acids remain attached to the gel layer.

7. The method of claim 5, wherein the contacting occurs under conditions wherein a uniform concentration of the nucleic acids contacts the portions of the continuous gel layer that coat the mask regions and the transparent regions.

8. The method of claim 5, wherein the continuous gel layer comprises a continuous polyacrylamide layer.

9. The method of claim 5, wherein the nucleic acids that attach to the portions of the gel layer that coat the mask regions have the same sequence.

10. The method of claim 5, wherein the nucleic acids that attach to the portions of the gel layer that coat the mask regions comprise a pair of primers.

12. A method of making an analyte array, comprising

(a) providing a surface comprising mask regions and transparent regions, the mask regions having a composition that blocks transmittance of radiation in a first part of the electromagnetic spectrum and transparent regions having a composition that transmits radiation in the first part of the electromagnetic spectrum, wherein a continuous gel layer coats the surface across the mask regions and the transparent regions;

(b) contacting the continuous gel layer with a fluid comprising a population of different analytes under conditions wherein analytes from the population become attached to the gel layer and wherein a first subpopulation of the analytes attach to portions of the gel layer that coat the mask regions and a second subpopulation of the analytes attach to portions of the gel layer that coat the transparent regions; and

(c) irradiating the surface with radiation in the first part of the electromagnetic spectrum, thereby selectively modifying the analytes of one of the subpopulations compared to the analytes of the other subpopulation,

wherein the surface of the mask regions comprise metal, thereby forming metal regions on the surface.

14. The method of claim 12 or 13, wherein step (c) comprises selectively modifying the analytes of the second subpopulation compared to the analytes of the first subpopulation.

15. The method of claim 14, wherein the selectively modifying comprises photo-chemically activating the analytes of the second subpopulation.

16. The method of claim 12 or 13, wherein step (c) comprises selectively modifying the analytes of the first subpopulation compared to the analytes of the second subpopulation.

17. The method of claim 16, wherein the selectively modifying comprises photo-chemically inactivating the analytes of the first population.

28. The method of claim 12 or 13, wherein the analytes comprise nucleic acids.

29. The method of claim 28, further comprising amplifying the nucleic acids on the portions of the gel layer that coat the mask regions.

30. The method of claim 29, further comprising removing the mask regions from the surface under conditions wherein the products of the amplifying of the nucleic acids remain attached to the gel layer.

31. The method of claim 29, wherein the contacting occurs under conditions wherein a uniform concentration of the nucleic acids contacts the portions of the continuous gel layer that coat the mask regions and the transparent regions.

32. The method of claim 29, wherein the continuous gel layer comprises a continuous polyacrylamide layer.

33. The method of claim 29, wherein the nucleic acids that attach to the portions of the gel layer that coat the mask regions have the same sequence.

34. The method of claim 29, wherein the nucleic acids that attach to the portions of the gel layer that coat the mask regions comprise a pair of primers.

13. A method of making an analyte array, comprising

(a) providing a surface comprising mask regions and transparent regions, the mask regions having a composition that blocks transmittance of radiation in a first part of the electromagnetic spectrum and transparent regions having a composition that transmits radiation in the first part of the electromagnetic spectrum, wherein a continuous gel layer coats the surface across the mask regions and the transparent regions;

(b) contacting the continuous gel layer with a fluid comprising a population of different analytes under conditions wherein analytes from the population become attached to the gel layer and wherein a first subpopulation of the analytes attach to portions of the gel layer that coat the mask regions and a second subpopulation of the analytes attach to portions of the gel layer that coat the transparent regions; and

(c) irradiating the surface with radiation in the first part of the electromagnetic spectrum, thereby selectively modifying the analytes of one of the subpopulations compared to the analytes of the other subpopulation,

wherein the surface is located in a flow-cell.

18. A method of making a nucleic acid array, comprising

(a) providing a surface comprising metal regions and interstitial regions, the interstitial regions having a composition that differs from the metal regions, wherein a continuous gel layer coats the surface across the metal regions and the interstitial regions;

(b) contacting the continuous gel layer with a fluid comprising analytes; and

(c) selectively modifying the portions of the gel layer that coat the interstitial regions compared to the portions of the gel layer that coat the metal regions,

thereby attaching the analytes to the gel layer, wherein a greater amount of the analytes from the fluid attach to portions of the gel layer that coat the metal regions than the amount of analytes that attach to portions of the gel layer that coat the interstitial regions.

21. The method of claim 18, wherein the analytes comprise nucleic acids.

22. The method of claim 21, wherein the contacting of the continuous gel layer with the fluid comprising the nucleic acids occurs prior to the modifying of the portions of the gel layer that coat the interstitial regions.

23. The method of claim 21, further comprising removing the metal regions from the surface under conditions wherein the nucleic acids remain attached to the gel layer on the surface.

24. The method of claim 23, wherein the metal regions are removed subsequent to the selectively modifying.

25. The method of claim 21, wherein the selectively modifying comprises selectively removing the nucleic acids that are attached to the portions of the gel layer that coat the interstitial regions compared to the nucleic acids that are attached to the portions of the gel layer that coat the metal regions.

26. The method of claim 21, wherein the selectively modifying comprises selectively inactivating the nucleic acids that are attached to the portions of the gel layer that coat the interstitial regions compared to the nucleic acids that are attached to the portions of the gel layer that coat the metal regions.

27. The method of claim 21, further comprising amplifying template nucleic acids on the portions of the gel layer that coat the metal regions.

19. A method of making a nucleic acid array, comprising

(a) providing a surface comprising metal regions and interstitial regions, the interstitial regions having a composition that differs from the metal regions, wherein a continuous gel layer coats the surface across the metal regions and the interstitial regions;

and

(b) contacting the continuous gel layer with a fluid comprising analytes under conditions wherein the analytes become attached to the gel layer and wherein a greater amount of the analytes from the fluid attach to portions of the gel layer that coat the metal regions than the amount of analytes that attach to portions of the gel layer that coat the interstitial regions,

wherein the analytes comprise nucleic acids,

further comprising removing the metal regions from the surface under conditions wherein the nucleic acids remain attached to the gel layer on the surface.

20. The method of claim 19, wherein the contacting occurs under conditions wherein a uniform concentration of the nucleic acids contacts the portions of the continuous gel layer that coat the metal regions and the interstitial regions.