Method of manufacturing a matrix for the detection of mismatches

• US 5,770,721 A
• Filed: 05/19/1995
• Issued: 06/23/1998
• Est. Priority Date: 08/11/1993
• Status: Expired due to Term

• Current Assignee: U Chicago Argonne, LLC
• Original Assignee: University of Chicago
• Inventors: Mirzabekov, Andrei Darievich, Ershov, Gennady Moiseevich
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): REES, DIANE
• Application Number:
  US08/411,794
• Time in Patent Office: 1,131 Days
• Field of Search: 435/6, 435/5, 435/91.1, 435/7.1-7.9, 435/7.72, 430/320, 430/311, 430/313, 430/325, 536/25.3, 536/24.3, 436/525, 436/528, 436/809, 427/2.22, 429/44, 429/25, 156/628, 156/632.1, 156/638.1, 156/643.1, 205/139, 205/149, 205/153, 205/159, 205/162, 216/43, 216/48, 216/51, 216/41, 216/55, 216/65, 216/66, 216/75, 216/94, 264/139, 264/176, 422/57, 422/61
• US Class Current: 536/25.3
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00531 essentially square
• CPC Class Codes: B01J 2219/00644 the porous medium being present in discrete locations, e.g. gel pads
• CPC Class Codes: B01J 2219/00646 the compounds being bound to beads immobilised on the solid supports
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof

Abstract

This method for preparing micromatrices consists in applying a specially-patterned intermediate layer of laser-absorbing substance on a solid support. The configuration of the sublayer fully corresponds to the topology of the manufactured matrix. The intermediate layer is further covered by a continuous layer of gel , the gel and the material of the support being transparent towards laser radiation. The gel layer is irradiated by a laser beam for a time needed to evaporate simultaneously the gel in the places immediately above the laser-absorbing sublayer and the sublayer itself. Oligonucleotides from a chosen set are then attached to the formed gel `cells`, one oligonucleotide to each cell.

This method is intended for use in biotechnology, specifically for deciphering the nucleotide sequence of DNA.

Claims

1. A method to prepare a patterned matrix for immobilizing oligonucleotides at distinct intervals on said matrix, comprising:

(a) precoating a solid support at distinct intervals with a sublayer of a substance to form a precoated matrix with a prescribed topology, wherein said solid support is virtually transparent to laser radiation and wherein said sublayer substance absorbs laser radiation;

(b) applying a continuous gel coat onto said precoated matrix wherein said continuous gel coat is also virtually transparent to laser radiation to form a gel coated matrix;

(c) irradiating said gel coated matrix with a laser beam so that upon irradiation those parts of the continuous gel coat which are immediately above said sublayer substance are evaporated together with said substance, wherein said evaporation forms a multiplicity of gel cells in the gel coated matrix which are spaced from one another by intervals forming a patterned matrix; and

(d) immobilizing oligonucleotides from a chosen set of oligonucleotides into the gel cells wherein one oligonucleotide is immobilized per gel cell.

2. The method of claim 1 wherein said sublayer substance is aluminum.

3. The method of claim 1 wherein said sublayer substance has a thickness directly proportional to the thickness of the gel coat.

4. The method of claim 2 wherein said sublayer substance has a thickness directly proportional to the thickness of the gel coat.