IP Library Granted Patent US 8,563,477
Granted Patent B2
US 8,563,477 · App. 13/548,558 · Granted Oct 22, 2013

Modified molecular arrays

Inventors: Mark Edward Brennan Smith (Walden, GB); Andrea Sabot (Walden, GB); Isabelle Marie Julia Rasalonjatovo (Walden, GB); Jean-Ernest Sohna Sohna (Walden, GB); Adrian Martin Horgan (Walden, GB); Harold Philip Swerdlow (Walden, GB)
Assignee: Illumina Cambridge Limited
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Quick Facts
Patent No.
US 8,563,477
App. No.
13/548,558
Granted
Oct 22, 2013
Kind
B2
Abstract

A composition including (a) a solid support having a surface; (b) a first plurality of nucleic acids immobilized on the surface, wherein the nucleic acids in the first plurality each include the P7 primer sequence (5′-CAAGCAGAAGACGGCATACGA-3′; SEQ ID NO: 3); and (c) a second plurality of nucleic acids immobilized on the surface, wherein the nucleic acids in the second plurality each include the P5 primer sequence (5′-AATGATACGGCGACCACCGA-3′; SEQ ID NO: 4).

Claims (34)

1. A composition comprising:

(a) a solid support comprising a surface;

(b) a first plurality of nucleic acids immobilized on the surface, wherein the nucleic acids in the first plurality each comprise the sequence 5′-CAAGCAGAAGACGGCATACGA-3′ (SEQ ID NO: 3); and

(c) a second plurality of nucleic acids immobilized on the surface, wherein the nucleic acids in the second plurality each comprise the sequence 5′-AATGATACGGCGACCACCGA-3′ (SEQ ID NO: 4).

2. The composition of claim 1 , wherein the nucleic acids are covalently attached to a hydrogel that is immobilized on the surface.

3. The composition of claim 2 , wherein the thickness of the hydrogel is less than 100 nm.

4. The composition of claim 2 , wherein the 5′ ends of the nucleic acids are covalently attached to the hydrogel.

5. The composition of claim 2 , wherein the hydrogel is the product of polymerizing a first comonomer with a second comonomer, wherein the first comonomer is selected from the group consisting of acrylamide, methacrylamide, hydroxyethyl methacrylate and N-vinyl pyrrolidinone, and wherein the second comonomer is selected from the group consisting of a functionalized acrylamide, acrylate, methacrylate and methacrylamide.

6. The composition of claim 2 , wherein the hydrogel is the product of polymerizing acrylamide with N-(5-bromoacetamidylpentyl)acrylamide (BRAPA).

7. The composition of claim 1 , wherein the nucleic acids are covalently attached to a polyelectrolyte multilayer that is immobilized on the surface.

8. The composition of claim 1 , wherein the nucleic acids are covalently attached directly to the surface.

9. The composition of claim 1 , wherein the nucleic acids in the first plurality each comprise the sequence 5′-CAAGCAGAAGACGGCATACGA (SEQ ID NO: 3)-template-TCGGTGGTCGCCGTATCATT-3′ (SEQ ID NO: 5), wherein the template comprises a nucleotide sequence that is longer than 300 nucleotides.

10. The composition of claim 1 , wherein the nucleic acids in the second plurality each comprise the sequence 5′-AATGATCGGCGACCACCGA (SEQ ID NO: 4)-template-TCGTATGCCGTCTTCTGCTTG (SEQ ID NO: 11)-3′, wherein the template comprises a nucleotide sequence that is longer than 300 nucleotides.

11. The composition of claim 1 , wherein the nucleic acids in the first plurality each comprise the sequence 5′-polyT-CAAGCAGAAGACGGCATACGA (SEQ ID NO: 3)-3′, wherein the polyT comprises from 2 to 10 thymine nucleotides.

12. The composition of claim 1 , wherein the nucleic acids in the second plurality each comprise the sequence 5′-polyT-AATGATACGGCGACCACCGA (SEQ ID NO: 4)-3′, wherein the polyT comprises from 2 to 10 thymine nucleotides.

13. The composition of claim 1 , wherein the composition comprises an array of nucleic acid clusters, wherein each of the clusters comprises nucleic acids in the first plurality and nucleic acids in the second plurality.

14. The composition of claim 13 , wherein each of the clusters comprises nucleic acids in the first plurality that are hybridized to nucleic acids in the second plurality.

15. The composition of claim 1 , wherein the solid support comprises a silica-based substrate, plastic, gold, titanium dioxide or silicon.

16. The composition of claim 1 , wherein the nucleic acids in the first plurality are hybridized to target nucleic acids comprising the sequence 5′-TCGGTGGTCGCCGTATCATT-3′ (SEQ ID NO: 5).

17. The composition of claim 1 , wherein the nucleic acids in the second plurality are hybridized to target nucleic acids comprising the sequence 5′-TCGTATGCCGTCTTCTGCTTG-3′ (SEQ ID NO: 11).

18. The composition of claim 1 , wherein the sequence 5′-CAAGCAGAAGACGGCATACGA-3′ (SEQ ID NO: 3) is at the 3′ end of the nucleic acids in the first plurality.

19. The composition of claim 1 , wherein the sequence 5′-AATGATACGGCGACCACCGA-3′ (SEQ ID NO: 4) is at the 3′ end of the nucleic acids in the second plurality.

20. The composition of claim 1 , wherein the nucleic acids in the first plurality further comprise a fluorescent label.

21. The composition of claim 20 , wherein the nucleic acids in the first plurality further comprise a 3′ blocking group.

22. The composition of claim 1 , wherein the nucleic acids in the second plurality further comprise a fluorescent label.

23. The composition of claim 22 , wherein the nucleic acids in the second plurality further comprise a 3′ blocking group.

24. The composition of claim 1 , wherein the surface comprises a polyacrylamide based polymer.

25. The composition of claim 1 , wherein the surface does not comprise silane.

26. The composition of claim 1 , wherein the surface comprises polymethylmethacrylate.

27. The composition of claim 1 , wherein the surface comprises a polymer comprising N-(5-bromoacetamidyl pentyl)acrylamide (BRAPA).

28. The composition of claim 1 , wherein the surface has a thickness of less than 50 nm.

29. The composition of claim 1 , wherein the solid support comprises glass.

30. The composition of claim 1 , wherein the solid support comprises a polyelectrolyte.

31. A method of determining the sequence of a polynucleic acid comprising the steps of contacting said polynucleic acid with the composition of claim 1 and thereafter determining the sequence of the polynucleic acid.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 10, 2012
From: SMITH, MARK E.; SABOT, ANDREA; RASOLONJATOVO, ISABELLE; HORGAN, ADRIAN M.; SWERDLOW, HAROLD P.
To: ILLUMINA CAMBRIDGE LIMITED
Reel/Frame 028929/0399 →
Priority Claims (2)
GB 0400253.1 · Jan 7, 2004 · national
EP 04254726 · Aug 5, 2004 · regional
Continuity (2)
Continuation 10585373
Related Publication 20130085084A1 · Apr 4, 2013