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Gold deposition detection method for gene chip

A detection method and gene chip technology, which are applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of limited kit development, high toxicity of silver staining reagents, strong light sensitivity, etc., and improve detection sensitivity. , Simple operation, high detection sensitivity

Active Publication Date: 2013-12-18
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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Problems solved by technology

However, due to the high toxicity and strong photosensitivity of silver staining reagents, it is difficult to preserve, which limits the development of kits and their development in grassroots units, hospitals and other units.

Method used

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  • Gold deposition detection method for gene chip
  • Gold deposition detection method for gene chip
  • Gold deposition detection method for gene chip

Examples

Experimental program
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Embodiment 1

[0035] Example 1: Comparison of the results of detection of Mycobacterium tuberculosis based on hydrogen peroxide reduction and gene chip gold deposition detection methods based on hydroxylamine reduction and silver staining

[0036] 1. Chip Preparation

[0037] Amino-modified probes (see Table 1 for the probe sequence) were dot-patterned on the surface of the aldehyde-modified chip using a chip spotter, and the dot-patterned chip was placed in a closed space with a relative humidity of 75% and a temperature of 30°C for 24 hours. ~48 hours.

[0038] Table 1 Nucleic acid probe sequence list

[0039]

[0040] Note: Biotin: biotin; NH2: amino, all sequences were synthesized by Takara

[0041] 2. Preparation of nano-gold composite probes

[0042] With 0.1mol / L K 2 CO 3 Adjust the pH value of the nano-gold solution (20 μl / ml) to pH 6, and add 1 / 10 volume of streptavidin solution (60 μg / ml) (the concentration is determined by the nano-gold protein stability experiment). Mix...

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Abstract

The invention provides a gold deposition detection method for a gene chip. The method is characterized by comprising the steps of carrying out sample application on an existing DNA sequence to a micro array to prepare the gene chip, previously decorating a biological macromolecule at the 5' end of nucleic acid to be detected, marking the other biological macromolecule matched with a decorative molecule on the nucleic acid to be detected on nanogold to establish a nanogold composite probe, then mixing the decorated nucleic acid to be detected, the marked nanogold composite probe and the gene chip, incubating, washing the unreacted nanogold composite proble and adding hydrogen peroxide gold to enhance reaction liquid observation. The gold deposition detection method for the gene chip has the advantages of being high in flexible, convenient to detect, simple to operate, short in consumed time, low in cost, visual in detection and low in relying degree on detecting and signal reading equipment.

Description

technical field [0001] The invention relates to the technical field of biochip detection, in particular to a gold deposition detection method for a gene chip. Background technique [0002] Microarrays are formed by using the intensive and parallel processing principles of microelectronic chip technology to orderly solidify a large number of biological samples with biological significance on silicon substrates or glass. Using microarrays, a large number of biological samples can be quickly obtained or processed. information. Microarray-based signal labeling usually uses fluorescent labeling detection methods, but the disadvantages of expensive fluorescent reagents, biomolecular autofluorescence interference, fluorescence attenuation, and expensive reading instruments also greatly limit its application in medical clinical diagnosis. ZL01113298.1 provides a simple, fast, and low-cost gene chip detection technology. This technology uses the naked eye or ordinary optical scanner...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 景奉香贾春平张冀申王文涛金庆辉赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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