Matrix arrays and methods for making same

• US 10,066,260 B2
• Filed: 03/13/2014
• Issued: 09/04/2018
• Est. Priority Date: 03/14/2013
• Status: Active Grant

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Light, David, Hinz, Wolfgang, Cicero, Ronald L., Inman, Christina E., Kenney, Paul M., Mastroianni, Alexander, Rozhkov, Roman, Wang, Yufang, Gray, Jeremy, Glazer, Marc, Gremyachinskiy, Dmitriy
• Primary Examiner(s): Bhat, Narayan
• Application Number:
  US14/774,662
• Publication Number:
  US 20160032371A1
• Time in Patent Office: 1,636 Days
• 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 1/6834 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 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2523/101 Crosslinking agents, e.g. psoralen
• CPC Class Codes: C12Q 2523/301 Sonication
• CPC Class Codes: C12Q 2523/32 Centrifugation
• CPC Class Codes: C12Q 2527/119 pH
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2527/156 Permeability
• CPC Class Codes: C12Q 2535/122 Massive parallel sequencing
• CPC Class Codes: C12Q 2547/107 Use of permeable barriers, e.g. waxes
• CPC Class Codes: C12Q 2563/159 Microreactors, e.g. emulsion PCR or sequencing, droplet PCR, microcapsules, i.e. non-liquid containers with a range of different permeability's for different reaction components
• CPC Class Codes: C12Q 2565/537 characterised by the capture oligonucleotide acting as a primer
• CPC Class Codes: C12Q 2565/629 being a microfluidic device

Abstract

A method of forming a polymer matrix array includes applying an aqueous solution into wells of a well array. The aqueous solution includes polymer precursors. The method further includes applying an immiscible fluid over the well array to isolate the aqueous solution within the wells of the well array and polymerizing the polymer precursors isolated in the wells of the well array to form the polymer matrix array. An apparatus includes a sensor array, a well array corresponding to the sensor array, and an array of polymer matrices disposed in the well array.

Claims

1. A method of forming a polymer matrix array, the method comprising:

flowing an aqueous solution over wells of a well array, the aqueous solution comprising polymer precursors, the aqueous solution entering the wells;

displacing the aqueous solution from over the wells by flowing an immiscible fluid over the well array to isolate the aqueous solution comprising the polymer precursors within the wells of the well array; and

polymerizing the polymer precursors in the aqueous solution isolated in the wells of the well array by the immiscible fluid to form the polymer matrix array wherein the wells of the well array overlie sensors of a sensor array, the wells corresponding to ion sensitive material of the sensors of the sensor array.

2. The method of claim 1, wherein the well has a characteristic diameter in a range of 0.1 micrometers to 2 micrometers.

3. The method of claim 1, wherein the well has a thickness in a range of 0.01 micrometers to 10 micrometers.

4. The method of claim 1, wherein a polymer matrix of the polymer matrix array is conformal with walls of a well of the well array.

5. The method of claim 1, wherein the polymer precursors include a radically polymerizable monomer.

6. The method of claim 5, wherein the radically polymerizable monomer includes a vinyl-based monomer.

7. The method of claim 5, wherein the radically polymerizable monomer includes acrylamide, vinyl acetate, hydroxyalkylmethacrylate, variations or derivatives thereof or any combination thereof.

8. The method of claim 1, wherein the polymer precursors include a crosslinker.

9. The method of claim 8, wherein the crosslinker is a bis-acrylamide.

10. The method of claim 1, wherein the polymer precursors include a surface active additive.

11. The method of claim 1, wherein the polymer precursors include an oligonucleotide functionalized acrylamide.

12. The method of claim 1, wherein the polymer precursors include N-(5- bromoacetamidylpentyl)acrylamide.

13. The method of claim 1, wherein applying the aqueous solution includes applying an emulsion including the aqueous solution as a dispersed phase.

14. The method of claim 1, further comprising spinning the well array following flowing the aqueous solution.

15. The method of claim 1, further comprising sonicating the well array following flowing the aqueous solution.

16. The method of claim 1, further comprising degassing the well array during flowing the aqueous solution.

17. The method of claim 1, further comprising wetting the well array with a hydrophilic fluid prior to flowing the aqueous solution.

18. The method of claim 1, wherein the immiscible fluid includes an initiator.

19. The method of claim 1, further comprising treating a surface within the well with a surface active agent prior to flowing the aqueous solution.