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Method of using gene chip technology to identify and detect 16 animal-derived compositions

An animal-derived component and gene chip technology, applied in the field of biology, can solve the problems of low detection efficiency and complex detection methods, and achieve the effects of high sensitivity, simple operation and strong specificity

Inactive Publication Date: 2012-10-31
宗卉
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] 1. The purpose of the present invention is to provide a series of highly efficient and specific oligonucleotide probes for the detection of animal-derived components for the above-mentioned problems of complex detection methods and low detection efficiency

Method used

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  • Method of using gene chip technology to identify and detect 16 animal-derived compositions
  • Method of using gene chip technology to identify and detect 16 animal-derived compositions
  • Method of using gene chip technology to identify and detect 16 animal-derived compositions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The preparation of embodiment 1 gene chip

[0043] 1. Design of primers and probes

[0044] According to the target DNA sequence homology analysis results, select each target specific DNA sequence, screen out all DNA fragments suitable for designing probes in the general primer amplification interval, use the primer and probe analysis software PrimerPremier 5.0 (this software can Various parameters of primers and probes are given, and primers and probes are effectively evaluated in many necessary aspects) to design and analyze to ensure the scientificity and practicability of the design. In order to ensure that all probes on the chip can get better hybridization effect at one hybridization temperature, the design of primers and probes should strictly follow the general design principles.

[0045]Submit the candidate sequences to GenBank for Blast comparison analysis. After screening, oligonucleotide sequences with theoretically better specificity are selected as detect...

Embodiment 2

[0053] Embodiment 2 gene chip detection method

[0054] 1. Materials and Instruments

[0055] 1. Materials

[0056] 1.1 Sample

[0057] Beef, goat, sheep, buffalo, pork, dog, rabbit, chicken, duck, goose, quail, turkey, ostrich and tilapia were purchased from a supermarket in Shenzhen, Guangdong, donkey and venison purchased from a supermarket in Hohhot, Inner Mongolia. These samples were all identified by species and verified by sequencing, and the results proved that they were all the species mentioned in their names.

[0058] 1.2 Main reagents

[0059] DMSO (Shanghai Sangon Bioengineering Co., Ltd.)

[0060] SDS (Shanghai Sangon Biological Engineering Co., Ltd.)

[0061] 1×TE (Shanghai Sangon Bioengineering Co., Ltd.)

[0062] NaBH 4 (Shanghai Sangon Bioengineering Co., Ltd.)

[0063] Ultrapure water (ddH 2 O)

[0064] Blank control: 50% DMSO

[0065] Spotting buffer: 50% DMSO

[0066] Washing solution after spotting: 0.2% SDS

[0067] Blocking solution after ...

Embodiment 3

[0100] Embodiment 3 specific detection

[0101] According to the above-mentioned test method, extract beef powder, goat meat powder, sheep meat powder, venison powder, buffalo meat powder, donkey meat powder, pork powder, dog meat powder, rabbit meat powder, chicken powder, duck meat powder, goose meat powder, quail meat powder, turkey meat powder, The DNA of 16 kinds of livestock and poultry, such as ostrich meat powder and tilapia meat powder, were subjected to chip hybridization experiments under the conditions of the optimal fluorescent labeling temperature combination to verify the specificity of the method. ( image 3 )

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Abstract

The invention discloses an oligonucleotide probe for detection of 16 animal-derived compositions, a gene chip preparation and operation method and a detection method of a gene chip made of the oligonucleotide probe for detection of 16 animal-derived compositions (including cattle, goats, sheep, deer, buffalos, donkeys, pigs, dogs, rabbits, chickens, ducks, gooses, quails, turkeys, ostriches and tilapias). Different animal-derived compositions in foods and feeds (including additives) containing animal-derived compositions can be rapidly and stably detected by the method.

Description

1. Technical field [0001] The invention relates to 16 animal oligonucleotide probes, a gene chip and a detection method thereof, and belongs to the technical field of biology. Background technique [0002] At present, in order to determine the true composition of food and feed, many methods for the identification of ingredients of animal origin have been developed. There have been research reports abroad on the ingredients derived from cattle, sheep, pigs, and chickens, and domestic research reports on cattle, sheep, pigs, chickens, donkeys, horses, and cervus. These methods include physical methods, chemical methods, immunology and molecular biology methods, but these methods have complex detection methods and low detection efficiency. A detection method can only target one target and cannot detect other target components. The current large-scale high-throughput detection requirements for feed and meat products, the present invention is a system of supplies and methods dev...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C40B40/06
Inventor 宗卉石丰运曹琛福
Owner 宗卉
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