Method and apparatus for performing large numbers of reactions using array assembly

• US 20030232382A1
• Filed: 06/23/2003
• Published: 12/18/2003
• Est. Priority Date: 10/08/1999
• Status: Abandoned Application

• Current Assignee: Francois Chatelain, Mark Berninger, Thomas M. Brennan
• Original Assignee: Francois Chatelain, Mark Berninger, Thomas M. Brennan
• Inventors: Chatelain, Francois, Berninger, Mark, Brennan, Thomas M.
• Application Number:
  US10/602,998
• Publication Number:
  US 20030232382A1
• Time in Patent Office: Days
• US Class Current: 435/6.18
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00635 by reactive plasma treatment
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2535/125 Allele specific primer extension
• CPC Class Codes: C12Q 2537/143 Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
• CPC Class Codes: C12Q 2565/515 : characterised by the interaction between or sequential use of two or more arrays
• CPC Class Codes: C12Q 2565/518 : characterised by the immobilisation of the nucleic acid sample or target
• CPC Class Codes: C12Q 2600/156 : Polymorphic or mutational markers
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof

Abstract

The present invention relates to a method and apparatus for performing a large number of reactions using array assembly. In particular, the present invention features a method and apparatus for performing a large number of chemical and biological reactions by bringing two arrays into close apposition and allowing reactants on the surfaces of two arrays to come into contact. The present invention is exemplified by performing a large number of polynucleotide amplification reactions using array assembly. In addition, the present invention features a method and apparatus for coupling the amplification of polynucleotides and the detection of sequence variations, expression levels, and functions thereof.

Claims

1. A solid support for performing a plurality of polynucleotide amplification reactions wherein a releasable primer for each amplification reaction is immobilized on an area of the surface of said solid support.

14. The solid support of claim 1 wherein said releasable primer is releasable by strand separation.

15. The solid support of claim 1 wherein said releasable primer is releasable by photolysis.

2. A system for amplifying a plurality of target nucleic acids, comprising:

(a) a first solid support wherein

(1) the surface of said first solid support comprises a plurality of derivatized areas;

(2) a primer for each target nucleic acid or a sequence complementary to said primer is immobilized on a derivatized area of the first solid support; and

(b) a second solid support wherein said second solid support comprises a plurality of wells and each well corresponds to a primer for each target nucleic acid.

4. The system according to any one of the claims 2-3, wherein said plurality of target nucleic acids are a plurality of subsequences of a polynucleotide.

5. The system of any one of claims 2-3, wherein said first or second solid support is glass.

6. The system according to any one of claims 2-3 wherein said immobilization is covalent.

7. The system according to any one of claims 2-3 wherein said immobilization is non-covalent.

8. The system according to any one of claims 2-3 wherein said primer is covalently linked to a cleavable moiety.

9. The system according to claim 8 wherein said cleavable moiety is cleavable by photolysis.

10. The system according to any one of the claims 2-3 wherein the surface of said first solid support comprises a plurality of hydrophilic areas.

11. The system according to any one of the claims 2-3 wherein the density of the derivatized areas on said first solid support is between about 10 to 10,000 per cm².

12. The system according to any one of the claims 2-3 wherein the size of said derivatized area on said first solid support is between about 10⁻

3 to 5 mm².

13. The system according to any one of the claims 2-3 wherein the number of derivatized areas on said first solid support is between about 10 to 500,000.

3. A system for amplifying a plurality of target nucleic acids, comprising:

(a) a first solid support wherein

(1) the surface of said first solid support comprises a plurality of derivatized areas;

(2) a forward primer for each target nucleic acid or a sequence complementary to said forward primer is immobilized on a derivatized area of the first solid support

(3) a reverse primer for each region of said target nucleic acid or a sequence complementary to said reverse primer is immobilized on another derivatized area of the first solid support;

(b) a second solid support wherein said second solid support comprises a plurality of wells and each well corresponds to the forward and reverse primers for each target nucleic acid.