Fixation of nucleotide derivatives to solid carrier

• US 20050019790A1
• Filed: 02/12/2004
• Published: 01/27/2005
• Est. Priority Date: 08/09/2000
• Status: Abandoned Application

• Current Assignee: Hiroshi Shinoki, Kosuke Tashiro, Satoru Kuhara, Shigeru Muta, Yoshihide Iwaki, Yoshihiko Makino
• Original Assignee: Hiroshi Shinoki, Kosuke Tashiro, Satoru Kuhara, Shigeru Muta, Yoshihide Iwaki, Yoshihiko Makino
• Inventors: Muta, Shigeru, Iwaki, Yoshihide, Kuhara, Satoru, Makino, Yoshihiko, Tashiro, Kosuke, Shinoki, Hiroshi
• Application Number:
  US10/777,585
• Publication Number:
  US 20050019790A1
• Time in Patent Office: Days
• US Class Current: 435/6
• 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 2219/00351 Means for dispensing and evacuation of reagents
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00529 DNA chips
• 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 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 60/14 for creating libraries

Abstract

A micro-array for analysis of DNA is prepared by the steps of spotting onto a solid carrier in its predetermined area in which plural reactive groups are fixed an aqueous solution which contains a thickening agent and probe molecules (e.g., DNA fragments) having a group reactive with the reactive group of the solid carrier to produce covalent bonding; spotting onto the solid carrier in a different area having the same reactive groups an aqueous solution (same or different); incubating the aqueous solution-spotted solid carrier to produce the covalent bondings; and washing the solid carrier with an aqueous medium to remove the thickening agent from the solid carrier. An electrostatic bonding can be utilized in place of the covalent bonding.

Claims

1. A process for preparing a micro-array for analysis of DNA which comprises the steps of:

spotting onto a solid carrier in a predetermined area thereof in which a number of reactive groups are fixed an aqueous solution which contains a thickening agent and probe molecules having a group reactive with the reactive groups of the carrier to produce covalent bonding, the thickening agent being incorporated in an amount to increase viscosity of the solution to a predetermined value and the probe molecules being selected from the group consisting of nucleic acid fragments, oligonucleotides and peptide nucleic acids;

spotting onto the solid carrier in an area other than the area in which the aqueous solution was spotted and in which the same reactive groups are present an aqueous solution which contains a thickening agent and probe molecules having a group reactive with the reactive groups of the carrier to produce covalent bonding, the thickening agent being incorporated in an amount to increase viscosity of the solution to a predetermined value and the probe molecules being selected from the group consisting of nucleic acid fragments, oligonucleotides and peptide nucleic acids;

incubating the solid carrier having the spotted aqueous solutions on the surface to cause reaction for producing the covalent bondings; and

washing the surface of the solid carrier with an aqueous medium to remove the thickening agent from the surface of the solid carrier.

2. The process of claim 1, wherein each of the aqueous solutions to be spotted onto the solid carrier has a viscosity of 2 to 50 mPa·

s.

3. The process of claim 1, wherein the thickening agent is a water-soluble polymer.

4. The process of claim 1, wherein the reactive group of the solid carrier is a vinylsulfonyl group and the reactive group of the probe molecule is an amino group.

5. The process of claim 4, wherein the vinylsulfonyl group is provided to the solid carrier by reacting a divinylsulfone compound with a amino group which has been previously placed on the solid carrier.

6. The process of claim 1, wherein the aqueous medium for washing the solid carrier contains a surface active agent.

7. The process of claim 1, wherein each of the aqueous solutions has a viscosity essentially identical to each other.

8. A micro-array for analysis of DNA which is prepared by claim 1.

16. A micro-array for analysis of DNA which is prepared by claim 1.

9. A process for preparing a micro-array for analysis of DNA which comprises the steps of:

spotting onto a solid carrier in a predetermined area thereof in which a number of groups electrically chargeable in an aqueous medium are fixed an aqueous solution which contains a thickening agent and probe molecules having a group electrically chargeable in an aqueous medium to produce electrostatic bonding with the electrically chargeable groups of the carrier, the thickening agent being incorporated in an amount to increase viscosity of the solution to a predetermined value and the probe molecules being selected from the group consisting of nucleic acid fragments, oligonucleotides and peptide nucleic acids;

spotting onto the solid carrier in an area other than the area in which the aqueous solution was spotted and in which the same electrically chargeable groups are present an aqueous solution which contains a thickening agent and probe molecules having a group electrically chargeable in an aqueous medium to produce electrostatic bonding with the electrically chargeable groups of the carrier, the thickening agent being incorporated in an amount to increase viscosity of the solution to a predetermined value and the probe molecules being selected from the group consisting of nucleic acid fragments, oligonucleotides and peptide nucleic acids;

incubating the solid carrier having the spotted aqueous solutions on the surface to cause reaction for producing the covalent bondings; and

washing the surface of the solid carrier with an aqueous medium to remove the thickening agent from the surface of the solid carrier.

10. The process of claim 9, wherein each of the aqueous solutions to be spotted onto the solid carrier has a viscosity of 2 to 50 mPa·

s.

11. The process of claim 9, wherein the thickening agent is a water-soluble polymer.

12. The process of claim 9, wherein the electrically chargeable group of the solid carrier is an amino group and the electrically chargeable group of the probe molecule is a phosphoric acid group.

13. The process of claim 12, wherein the amino group is provided to the solid carrier by treating the solid carrier with an aminosilane coupling agent or a polycation compound.

14. The process of claim 9, wherein the aqueous medium for washing the solid carrier contains a surface active agent.

15. The process of claim 9, wherein each of the aqueous solutions has a viscosity essentially identical to each other.