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Method for producing a biomolecule assay chip

Foreign code F110003679
File No. AF12-01EP
Posted date Jul 1, 2011
Country EPO
Application number 07827976
Gazette No. 2090888
Gazette No. 2090888
Date of filing Nov 1, 2007
Gazette Date Aug 19, 2009
Gazette Date Dec 23, 2015
International application number JP2007001197
International publication number WO2008053598
Date of international filing Nov 1, 2007
Date of international publication May 8, 2008
Priority data
  • 2007JP001197 (Nov 1, 2007) WO
  • P2006-297267 (Nov 1, 2006) JP
Title Method for producing a biomolecule assay chip
Abstract It is an object of the present invention to provide a method for producing a biomolecule assay chip using a microreactor technique, and a chip produced by the method.
The present invention provides: a method for producing a chip on which biomolecules are immobilized in an aligned state, which comprises (a) a step of producing a substrate 1 on which a plurality of biomolecules 1 of a single type are immobilized in an aligned state, (b) a step of adding reaction reagents for synthesizing biomolecules 2 to microreactors on a microreactor chip comprising the microreactors at positions overlapping with the sequence positions of the biomolecules 1 immobilized on the substrate 1 produced in the step (a), (c) a step of closely attaching the microreactor chip to the substrate 1 so that the reaction reagents for synthesizing the biomolecules 2 are allowed to come into contact with the biomolecules 1, so as to synthesize the biomolecules 2 in the microreactors, and (d) superposing the microreactor chip on a substrate 2 after completion of the step (c) so as to bind the biomolecules 2 onto the substrate 2; and a chip produced by the aforementioned method.
Scope of claims [claim1]
1. A method for producing a chip on which protein-nucleic acid complexes are immobilized in an aligned state, which comprises the following steps (a) to (e): (a) a step of producing a substrate 1 on which a plurality of DNA are immobilized in an aligned state; (b) a step of filling microreactors on a microreactor chip comprising the microreactors at positions overlapping with the sequence positions of DNA immobilized on the substrate 1 produced in the step (a), with reaction reagents for synthesizingmRNA; (c) a step of closely attaching the microreactor chip to the substrate 1 so that the reaction reagents for synthesizing mRNA are allowed to come into contact with DNA, and then carrying out a reaction of synthesizing mRNA in the microreactors; (d) superposing the microreactor chip on a substrate 2 so that the reaction solutions contained in the microreactors on the microreactor chip are allowed to come into contact with the substrate 2 after completion of the step (c), so as to immobilize mRNA on the substrate 2; and (e)immersing the chip with aligned mRNA on substrate 2 in a solution containing a cell-free translation system for synthesizing the protein-nucleic acid complex.
[claim2]
2. The method according to claim 1, wherein the step of producing "a substrate 1 on which a plurality of DNA are immobilized in an aligned state" described in the step (a) according to claim 1 further comprises the following steps (a) to (c): (a) a step of diluting mixed solutions of DNA and reaction reagents for amplifying DNA, and then filling the microreactors on the microreactor chip with the diluted solutions, so that a single molecule or less of DNA can be present therein in a probability distribution manner; (b) a step of carrying out a reaction of amplifying DNA; and (c) a step of superposing the microreactor chip on the substrate 1 so that the reaction solutions contained in the microreactors on the microreactor chip are allowed to come into contact with the substrate 1, so as to immobilize the amplified DNA on the substrate.
[claim3]
3. The method according to claim 2, wherein the reaction of amplifying DNA is a polymerase chain reaction.
[claim4]
4. The method according to any one of claims 1 to 3, wherein the DNA are those to which linker DNA ligates.
[claim5]
5. The method according to claim 4, wherein puromycin binds to the linker DNA.
[claim6]
6. The method according to any one of claims 1 to 5, wherein it comprises immobilizing avidin on the substrate 1 and thereby biotinylating DNA.
[claim7]
7. The method according to any one of claims 1 to 6, wherein the mRNA is immobilized on the substrate 2 via the puromycin-binding linker DNA.
  • Applicant
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • Inventor
  • NEMOTO NAOTO
  • ICHIKI TAKANORI
  • BIYANI MANISH
IPC(International Patent Classification)
Reference ( R and D project ) CREST Establishment of Innovative Manufacturing Technology Based on Nanoscience AREA
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