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Hybridization method as well as hybridization microarray and hybridization kit

A microarray, biopolymer technology, applied in chemical instruments and methods, biochemical equipment and methods, biological testing, etc., can solve the problems of pollution, incomplete filling of the solution, residual air, etc.

Inactive Publication Date: 2007-03-07
KURASHIKI BOSEKI KK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the volume of the hybridization solution may change with the composition of the wetting agent, resulting in contamination of both samples on the slide
[0008] In the second method of the prior art, the separable container is not completely filled with solution and air is often trapped in the container, leading to incorrect hybridization

Method used

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  • Hybridization method as well as hybridization microarray and hybridization kit
  • Hybridization method as well as hybridization microarray and hybridization kit
  • Hybridization method as well as hybridization microarray and hybridization kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1 × 10 was prepared with RNeasy Mini kit (Qiagen) 6 10 μg total RNA from HeLa cells. Cy3-labeled HeLa cDNA was prepared as a sample biopolymer from 10 μg of HeLa cell total RNA prepared using Labelstar Array Kit (Qiagen) and Cy3-dUTP (Amersham Biosciences). The amount of the sample biopolymer was adjusted to 120 μL with a 5×SSC solution containing 0.5% SDS (the solute molarity of the solution containing the sample biopolymer was 1.67 mol / L).

[0044] The solution containing the sample biopolymer prepared by the above method is dropped onto the hydrophilic area of ​​the microarray, the surface of the microarray has 24 hydrophilic areas and hydrophobic areas around these hydrophilic areas, and each hydrophilic area is immobilized with Four kinds of probe biopolymers are respectively 70-mers of glyceraldehyde-3-phosphate dehydrogenase, endonuclease G, pU 18 and lambda phage DNA with base sequences shown in sequences 1-4 in the sequence table Probes (synthesized in a DNA ...

Embodiment 2

[0048] Hybridization was carried out similarly to Example 1, but using 600 μL of 4.5×SSC containing 0.5% SDS (the solute molar concentration of the wetting agent was 1.50 mol / L, and the difference between the solute molar concentration of the solution containing the sample biopolymer and it was -10 %) as a wetting agent. When the amount of the solution containing the sample biopolymer was measured after hybridization, it was found that the amount increased by 4.9 μL and the liquid change was 4.1% compared to when it was added dropwise. Therefore, hybridization can be accomplished by bringing the solution containing the sample biopolymer into contact only with the slide glass immobilized with the probe biopolymer without increasing the volume of the solution containing the sample biopolymer and overflowing the hydrophilic region.

Embodiment 3

[0050] Hybridization was carried out similarly to Example 1, but using 600 μL of 5.4×SSC containing 0.5% SDS (the solute molarity of the wetting agent was 1.80 mol / L, and the difference between the solute molarity of the solution containing the sample biopolymer was +8 %) as a wetting agent. When the amount of the solution containing the sample biopolymer was measured after hybridization, it was found that the amount decreased by 12.0 μL and the liquid change was 10.0% compared to when it was added dropwise. Therefore, hybridization can be accomplished by bringing the solution containing the sample biopolymer into contact only with the slide glass immobilized with the probe biopolymer without drying the solution containing the sample biopolymer on the microarray.

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Abstract

A method of simply and reliably hybridizing a sample biopolymer and probe DNA with each other without using a glass coverslip and a closed vessel by hybridizing the sample biopolymer and a probe biopolymer with each other in a state that a solution containing the sample biopolymer is in contact with only a slide glass to which the probe biopolymer is immobilized in a closed vessel containing a solution having the same vapor pressure as the solution containing the sample biopolymer is provided. A microarray and a kit for hybridization employed for the present invention are also provided.

Description

technical field [0001] The present invention relates to a hybridization method for simultaneously hybridizing multiple sample biopolymers and probe biopolymers, a hybridization microarray and a hybridization kit. Background technique [0002] The method of hybridizing a nucleic acid or protein probe of known sequence to sample DNA (usually a sample biopolymer) labeled with a fluorescent substance is commonly used to identify / isolate molecules in vivo, especially for detection of target DNA or genetic DNA presence / absence. More specifically, the method is performed using a DNA chip (usually a microarray) formed by immobilizing probe DNA (usually a probe biopolymer) onto a glass slide. [0003] In order to effectively hybridize sample DNA and probe DNA in a small amount of sample DNA solution, the following two types of methods are generally used: In the first method, a solution containing sample DNA labeled with a fluorescent substance is dropped onto DNA is placed on a gla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68B01L3/00G01N33/53C12M1/00C12N15/09G01N33/566G01N37/00
CPCC12Q1/6837C12Q2527/109C12Q2527/137C12Q1/6876
Inventor 宮川功砂川义彦森下笃
Owner KURASHIKI BOSEKI KK