US 11,142,759 B2Granted

Method for selecting and amplifying polynucleotides

AssigneeIllumina, Inc.

InventorsAndrea Sabot, Roberto Rigatti, Min-Jui Richard Shen

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Abstract

The invention provides methods for controlling the density of different molecular species on the surface of a solid support. A first mixture of different molecular species is attached to a solid support under conditions to attach each species at a desired density, thereby producing a derivatized support having attached capture molecules. The derivatized support is treated with a second mixture of different molecular species, wherein different molecular species in the second mixture bind specifically to the different capture molecules attached to the solid support. One or more of the capture molecules can be reversibly modified such that the capture molecules have a different activity before and after the second mixture of molecular species are attached. In particular embodiments, the different molecular species are nucleic acids that are reversibly modified to have different activity in an amplification reaction.

Claims19

Dependent claims

A method of selecting and amplifying polynucleotides on a solid support, comprising:

(a) providing a nucleic acid sample comprising a plurality of template polynucleotides;
(b) providing a plurality of oligonucleotides immobilised on a solid support wherein the plurality of oligonucleotides comprises:
(i) a plurality of capture oligonucleotides each comprising a different sequence capable of hybridising to a selected region of the nucleic acid sample, and
(ii) a plurality of amplification oligonucleotides comprising blocked amplification oligonucleotides, wherein the capture oligonucleotides are immobilised at a density of at least 100 fold lower than the amplification oligonucleotides;
(c) applying the template polynucleotides to the solid support under conditions such that the template polynucleotides selectively hybridise to the capture oligonucleotides;
(d) extending the capture oligonucleotides to generate extension products complementary to the template polynucleotides;
(e) denaturing the template polynucleotides from the extension products;
(f) deblocking the blocked amplification oligonucleotides, and
(g) amplifying the extension products, wherein the amplifying comprises annealing the amplification oligonucleotides to the extension products.

6 more elements

Dependencies are inferred from the claim text.

Facts

Application: 16/786,198Filed 10 Feb 2020 · continuation
Priority: 10 Mar 2008
Published: US 2020/0239875 A130 Jul 2020
Granted: 12 Oct 2021
Term ends: Not estimatedThe source records don’t support an estimate.

Classification9 codes

C12N 15/1068
Isolating an individual clone by screening libraries: template mediated chemical library synthesis, e.g. chemical and enzymatical DNA-templated…
C40B 40/06
Libraries containing only organic compounds: libraries containing nucleotides or polynucleotides, or derivatives thereof
C12Q 1/6806
Involving nucleic acids: preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
C12Q 1/6837
Enzymatic or biochemical coupling of nucleic acids to a solid phase: using probe arrays or probe chips

Family

Application61/035,254Us provisional application · filed 10 Mar 2008
Application12/395,299Parent of this continuation-in-part · filed 27 Feb 2009
PCT/US2009/054945Parent of this continuation · filed 25 Aug 2009
Application14/611,991Parent of this continuation · filed 2 Feb 2015
US 8,999,642 B2Parent of this continuation · filed 7 Apr 2015
Application15/489,549Parent of this continuation · filed 17 Apr 2017
US 9,624,489 B2Parent of this continuation · filed 18 Apr 2017
US 2020/0239875 A1Pre-grant publication · published 30 Jul 2020
US 10,597,653 B2Parent of this continuation
Application13/387,078Parent of this continuation
Application13/387,078Parent of this continuation-in-part
US 2009/0226975 A1Published application
US 2012/0122737 A1Published application
US 2015/0148263 A1Published application
US 2017/0283794 A1Published application
US 11,142,759 B2This patent

From the US filing history. Foreign family members aren’t listed.