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Polymer coatings

A polymer coating technology, applied in the fields of chemistry, biology and material science, can solve the problem of inconvenient coating beads

Active Publication Date: 2015-04-08
ILLUMINA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Polymer coatings available on planar array surfaces may be inconvenient for coating beads due to aggregation induced by coating and the need to store beads in aqueous buffer after coating
This leads to some limitations in commercial applications

Method used

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[0257] In some embodiments of the above-described methods of making a polymeric coating affixed to a substrate surface, said polymeric coating is dissolved in an aqueous solution prior to covalently binding to said functional groups of said surface. In some embodiments, the matrix is ​​a bead.

[0258] A second aspect of the methods disclosed herein relates to methods for preparing polynucleotide arrays. In such an embodiment, the method may comprise the step of: (a) coating a plurality of first oligonucleotides and a plurality of second oligonucleotides with a polymer coating present on the surface of any one of the substrates described herein. Reactive sites on a polymer coating prepared by or by any of the methods described herein for immobilizing a polymer coating to a substrate surface; (b) said plurality of contacting a first oligonucleotide with a template to be amplified; and (c) amplifying the template using the first oligonucleotide and the second oligonucleotide, the...

Embodiment 1

[0303] Preparation of PAZAM

[0304] overview

[0305] Unless otherwise stated, all reactions were performed under nitrogen or argon atmosphere and starting materials were obtained from suppliers (Aldrich Chemical Company, Fisher Scientific, Dow) and used as received without further purification. All reaction temperatures reported indicate the temperature of the bath / air in contact with the reaction vessel. Anhydrous solvents are not required.

[0306] Recorded as deuterium oxide (99.9 at. %D for LC NMR) on a Bruker Avance 400 MHz instrument 1 H-NMR spectrum and 13 C-NMR spectrum. Chemical shifts are expressed in parts per million (ppm, δ) downfield from tetramethylsilane (TMS) and are referenced to the indicated solvent as internal standard.

[0307] GPC analysis was performed by Smithers Rapra Technology Limited using the following chromatographic conditions:

[0308] Instrumentation: Malvern / Viscotek Triple Detector Array TDA301, combined pump and autosampler.

...

Embodiment 2

[0347] Preparation of PAZAM derivatives

[0348] The synthesis of PAZAM derivatives is shown in Scheme 2. First, BRAPA (1c) was reacted with t-Boc protected hydroxylamine (2a) to form intermediate (2b), which was treated with dichloroacetic acid to form the oxoamine derivative of PAZAM (2c). 2c can then be grafted with aldehyde functionalized oligonucleotides to form 2d.

[0349] Scheme 2. Synthesis of oxoamine derivatives of PAZAM

[0350]

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Abstract

The present disclosure relates to polymer coatings covalently attached to the surface of a substrate and the preparation of the polymer coatings, such as poly(N-(5-azidoacetamidylpentyl)acrylamide-co-acrylamide) (PAZAM), in the formation and manipulation of substrates, such as molecular arrays and flow cells. The present disclosure also relates to methods of preparing a substrate surface by using beads coated with a covalently attached polymer, such as PAZAM, and the method of determining a nucleotide sequence of a polynucleotide attached to a substrate surface described herein.

Description

[0001] Cross References to Related Applications [0002] This application claims U.S. Application No. 61 / 657,508, filed June 8, 2012; U.S. Application No. 61 / 753,833, filed January 17, 2013; and U.S. Application No. 61 / 753,833, filed March 4, 2013 13 / 784,368, all of which are hereby incorporated by reference in their entirety. technical field [0003] In general, the present invention relates to the fields of chemistry, biology and materials science. More specifically, the present invention relates to polymer coatings covalently attached to the surface of substrates for detection and / or analysis of molecules such as nucleic acids and proteins. Background of the invention [0004] Polymer coated substrates are used in many technical applications. For example, implantable medical devices can be coated with biologically inert polymers. In another example, the polymer-coated matrix is ​​used to prepare and / or analyze biomolecules. Molecular analysis, such as certain nucleic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09D133/26
CPCC09D135/00B05D3/06C09D133/26B05D3/065Y10T428/24802Y10T428/24917Y10T428/24926Y10T428/2998Y10T428/31536Y10T428/31935C12Q1/6806C12Q1/6874
Inventor 韦恩·N·乔治丹尼尔·布拉顿安德鲁·A·布朗任洪吉赖安·C·史密斯
Owner ILLUMINA INC
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