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Method for extracting animal muscular tissue DNA efficiently and safely

A technology of animal muscle and animal tissue, which is applied in the field of molecular biology, can solve the problems of multiple extraction steps, damage, and time-consuming, and achieve the effects of simple process, improved detection efficiency, and time-saving

Pending Publication Date: 2016-10-05
TIANJIN INSTITUE OF QUALITY STANDARD & TESTING OF AGRICULTUAL PRODS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to overcome the shortcomings of traditional sample DNA extraction steps, such as many, complicated, time-consuming, and the use of a large amount of organic solvents to cause damage to experimenters, and discloses a method for efficiently and safely extracting DNA from animal muscle tissue

Method used

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  • Method for extracting animal muscular tissue DNA efficiently and safely
  • Method for extracting animal muscular tissue DNA efficiently and safely
  • Method for extracting animal muscular tissue DNA efficiently and safely

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Taking fresh beef as a sample, the method of the present invention is used to rapidly extract DNA. The preparation method is as follows:

[0065] (1) Take the muscle tissue sample of the animal to be tested, wash it with water, remove the connective tissue and fat in the animal tissue, cut it into small pieces of about 200 mg, put it into a mortar pre-cooled with liquid nitrogen, and then slowly Add liquid nitrogen into the mortar and grind quickly until the sample becomes a fine powder, take 100mg and add it to a 2mL sterilized centrifuge tube;

[0066] (2) Add 500 μL of SDS lysate and 6 μL of proteinase K, bathe in water at 65°C for 60 minutes, and mix by inverting constantly;

[0067] (3) After the water bath, centrifuge at 13200rpm / min for 5min; carefully transfer the upper aqueous phase into a new centrifuge tube, add 0.8 times isopropanol; transfer the mixed liquid into the adsorption column, centrifuge at 13200rpm / min 30s, discard the waste liquid;

[0068] (4...

Embodiment 2

[0081] Taking fresh pork as a sample, the method of the present invention is used to rapidly extract DNA. The preparation method is as follows:

[0082] (1) Take the muscle tissue sample of the animal to be tested, wash it with water, remove the connective tissue and fat in the animal tissue, cut it into small pieces of about 200 mg, put it into a mortar pre-cooled with liquid nitrogen, and then slowly Add liquid nitrogen into the mortar and grind quickly until the sample becomes a fine powder, take 100mg and add it to a 2mL sterilized centrifuge tube;

[0083] (2) Add 500 μL of SDS lysate and 6 μL of proteinase K, bathe in water at 65°C for 60 minutes, and mix by inverting constantly;

[0084] (3) After the water bath, centrifuge at 13200rpm / min for 5min; carefully transfer the upper aqueous phase into a new centrifuge tube, add 0.8 times isopropanol; transfer the mixed liquid into the adsorption column, centrifuge at 13200rpm / min 30s, discard the waste liquid;

[0085] (4...

Embodiment 3

[0098] Taking fresh chicken as a sample, the method of the invention is used to rapidly extract DNA. The preparation method is as follows:

[0099] (1) Take the muscle tissue sample of the animal to be tested, wash it with water, remove the connective tissue and fat in the animal tissue, cut it into small pieces of about 200 mg, put it into a mortar pre-cooled with liquid nitrogen, and then slowly Add liquid nitrogen into the mortar and grind quickly until the sample becomes a fine powder, take 100mg and add it to a 2mL sterilized centrifuge tube;

[0100] (2) Add 500 μL of SDS lysate and 6 μL of proteinase K, bathe in water at 65°C for 60 minutes, and mix by inverting constantly;

[0101] (3) After the water bath, centrifuge at 13200rpm / min for 5min; carefully transfer the upper aqueous phase into a new centrifuge tube, add 0.8 times isopropanol; transfer the mixed liquid into the adsorption column, centrifuge at 13200rpm / min 30s, discard the waste liquid;

[0102] (4) Add 7...

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Abstract

The invention discloses a method for extracting animal muscular tissue DNA easily, conveniently, safely and fast. The method comprises the steps that after pork, mutton, beef and chicken are washed with clean water, connective tissue and fat in animal tissue are removed; the animal tissue is shorn into small blocks of 200 mg, and the small blocks are placed in liquid nitrogen and fast ground till powder is obtained; 100 mg of the powder is taken to be placed in a 2-mL centrifuge tube, and the DNA is extracted through SDS splitting decomposition, isopropanol precipitating and adsorption column purification and can be directly used for PCR amplification. The DNA extracting method is adopted, and the whole process of DNA extraction of a sample can be completed within 2 hours. The quality standard of the extracted DNA meets the requirement of subsequent PCR amplification, meanwhile, the harm of various organic reagents to experiment staff in the extraction process of a traditional SDS method or guanidine thiocyanate method is avoided, and time is greatly shortened; the method is more suitable for achieving the aim of efficiently and fast extracting sample DNA in the animal source ingredient identification process, and has the advantages of being efficient, fast, low in cost, easy and convenient to operate, free of toxicity, safe and the like.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, relates to a preparation method of template DNA, and is an efficient, fast, low-cost, easy-to-operate, non-toxic and safe animal muscle tissue DNA extraction method. Background technique [0002] In recent years, with the prevalence of some diseases such as mad cow disease, sheep scrapie, foot-and-mouth disease, and bird flu in Europe and other countries in the world, as well as the widespread occurrence of adulteration and fraud of some animal-source foods, the animal Source identification has become an issue of great concern. With the development of biotechnology, DNA-based PCR technology is widely used to identify animal origin in food. Because the PCR method is relatively simple, with high specificity and sensitivity, this technology has been combined with other technologies at home and abroad. A variety of methods for identifying food animal-derived components have been establishe...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 赵新王永兰青阔陈锐朱珠
Owner TIANJIN INSTITUE OF QUALITY STANDARD & TESTING OF AGRICULTUAL PRODS
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