Patterned flow-cells useful for nucleic acid analysis

Patent metadata

Field	Value
Publication number	US-10787698-B2
Application number	US-201414304556-A
Country	US
Kind code	B2
Filing date	Jun 13, 2014
Priority date	Jun 9, 2011
Publication date	Sep 29, 2020
Grant date	Sep 29, 2020

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.

First claim

What is claimed is:

1. A method of making a nucleic acid array, comprising:
(a) providing a surface having mask regions and transparent regions, the transparent regions having a composition that differs from the mask regions;
(b) contacting the surface with a polymerizable material;
(c) polymerizing the polymerizable material to form a continuous gel layer on the surface, wherein the portions of the gel layer that coat the mask regions have greater mass than portions of the gel layer that coat the transparent regions;
(d) contacting the continuous gel layer with a fluid comprising nucleic acids;
(e) selectively and covalently attaching the nucleic acids to the gel layer, wherein the nucleic acids from the fluid that are attached to portions of the gel layer that coat the mask regions are at a greater density than the nucleic acids that are attached to portions of the gel layer that coat the transparent regions; and
(f) selectively photochemically cleaving the nucleic acids from the gel layer in the transparent regions.

2. The method of claim 1 , wherein the nucleic acids comprise primer sequences.

3. The method of claim 2 , further comprising hybridizing template nucleic acids to the nucleic acids attached to the gel layer.

4. The method of claim 1 , wherein the mask regions comprise metal regions.

5. The method of claim 1 , wherein the transparent regions on the surface form interstitial regions that completely surround the mask regions on the surface.

6. The method of claim 1 , 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.

7. The method of claim 6 , wherein the attaching occurs while the 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 1 , wherein the surface is located in a flowcell.

9. The method of claim 1 , wherein the continuous gel layer comprises agarose, gelatin, or a polyacrylamide polymer.

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

11. The method of claim 1 , wherein the nucleic acids comprise a photocleavable linker.

12. The method of claim 1 , wherein the gel material is polymerized silane-free acrylamide (SFA) and is formed by polymerization of silane-free acrylamide and N—(S-bromoacetamidylpentyl)acrylamide (BRAPA).

13. The method of claim 1 , wherein the polymerizing is accomplished by way of a temperature-dependent change in state.

14. The method of claim 1 , wherein the polymerizable material is acrylamide, methacrylamide, hydroxyethyl methacrylate, or N-vinyl pyrrolidinone, or derivatives or co-polymers thereof.

15. The method of claim 1 , wherein selectively photochemically cleaving the nucleic acids from the gel layer in the transparent regions involves:
positioning the surface over a light source; and
turning on the light source, whereby the mask regions mask light from for the nucleic acids attached to the gel layer that coat the mask regions and the nucleic acids attached to the gel layer that coat the interstitial regions are exposed to the light.

16. A method of making a nucleic acid array, comprising:
(a) providing a surface having mask regions and transparent regions, the transparent regions having a composition that differs from the mask regions;
(b) contacting the surface with a polymerizable material;
(c) polymerizing the polymerizable material to form a continuous gel layer on the surface, wherein the portions of the gel layer that coat the mask regions have greater mass than portions of the gel layer that coat the transparent regions;
(d) contacting the continuous gel layer with a fluid comprising nucleic acids;
(e) selectively and covalently attaching the nucleic acids to the gel layer, wherein the nucleic acids from the fluid that are attached to portions of the gel layer that coat the mask regions are at a greater density than the nucleic acids that are attached to portions of the gel layer that coat the transparent regions; and
(f) selectively inactivating the nucleic acids that are attached to the portions of the gel layer that coat the transparent regions compared to the nucleic acids that are attached to the portions of the gel layer that coat the mask regions.

17. The method of claim 3 , further comprising amplifying the template nucleic acids on the gel layer.

18. The method of claim 17 , wherein the amplifying comprises bridge amplifying to form nucleic acid clusters attached to the gel layer.

19. The method of claim 16 , wherein selectively inactivating the nucleic acids that are attached to the portions of the gel layer that coat the transparent regions involves exposing the nucleic acids that are attached to the portions of the gel layer that coat the transparent regions to photolysis or laser ablation, and wherein the mask regions mask the nucleic acids attached to the gel layer that coat the mask regions from the photolysis or laser ablation.

20. The method of claim 16 , wherein the mask regions comprise metal regions, and wherein the transparent regions comprise glass, plastic, or silica.

21. The method of claim 16 , wherein the continuous gel layer comprises agarose, gelatin, or a polyacrylamide polymer.

22. The method of claim 16 , wherein the polymerizable material is acrylamide, methacrylamide, hydroxyethyl methacrylate, or N-vinyl pyrrolidinone, or derivatives or co-polymers thereof.

23. The method of claim 16 , wherein the nucleic acids comprise primer sequences.

24. The method of claim 16 , wherein the transparent regions on the surface form interstitial regions that completely surround the mask regions on the surface.

Assignees

• Illumina Inc

Inventors

• Lin Shengrong
• Wu Yir-Shyuan
• Gunderson Kevin
• Moon John A

Classifications

• G01N27/447 Primary

  using electrophoresis · CPC title

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• C12Q1/6837

  using probe arrays or probe chips (C12Q1/6874 takes precedence) · CPC title