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Method for separating residual DNA from soy sauce, soybean milk and tomato ketchup

A technology for ketchup and soy sauce, which is applied in the field of DNA separation and achieves the effects of low cost, simple and effective method, high sensitivity and high detection rate

Inactive Publication Date: 2005-08-31
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If it is possible to extract high-quality trace amounts of DNA suitable for PCR templates from the tested samples, especially from those deeply processed or refined foods (soy sauce, salad oil, soybean milk, tomato sauce, biscuits, etc.) DNA remains a thorny issue in GMO safety testing

Method used

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  • Method for separating residual DNA from soy sauce, soybean milk and tomato ketchup
  • Method for separating residual DNA from soy sauce, soybean milk and tomato ketchup
  • Method for separating residual DNA from soy sauce, soybean milk and tomato ketchup

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Embodiment 1

[0023] The reagents used in this example are all promega products except that chloroform, ethanol, isoamyl alcohol and isopropanol are domestic reagents.

[0024] (1) CTAB method to extract residual DNA

[0025] Take 50ml of soy sauce, soy milk, and tomato paste, centrifuge at 12,000rpm for 10min, discard the supernatant, and save the precipitate for DNA extraction; or heat at 80°C to evaporate water and take dry matter for DNA extraction; or freeze-dry and take dry The material was used for DNA extraction. The powdered dry matter was added to 400 μl of pre-cooled cetyltrimethylammonium bromide (CTAB) extraction buffer I (50 mmol / L Tris-Cl, 10 mmol / L EDTA, 800 mmol / L NaCl, pH 8.0). Add 500 μl 65°C preheated CTAB extraction buffer II (2% CTAB, 50mmol / LTris-Cl, 10mmol / L EDTA, 800mmol / L NaCl, pH8.0), 1 μl 2-mercaptoethanol, mix well, and incubate at 65°C 30-90min, during which, gently invert and mix from time to time. After cooling to room temperature, add 450 μl of chloroform...

Embodiment 2

[0060] (1) CTAB method to extract residual DNA

[0061] Take 50ml of soy sauce, soy milk, and tomato paste, centrifuge at 12,000rpm for 10min, discard the supernatant, and save the precipitate for DNA extraction; or heat at 80°C to evaporate water and take dry matter for DNA extraction; or freeze-dry and take dry The material was used for DNA extraction. The powdered dry matter was added to 400 μl of pre-cooled CTAB extraction buffer I (50 mmol / L Tris-Cl, 10 mmol / L EDTA, 800 mmol / L NaCl, pH 8.0). Add 500 μl 65°C preheated CTAB extraction buffer II (2% CTAB, 50mmol / L Tris-Cl, 10mmol / L EDTA, 800mmol / L NaCl, pH 8.0), 1 μl 2-mercaptoethanol, mix well, 65°C Keep warm for 30-90min, during which time, gently invert and mix well. After cooling to room temperature, add 450 μl of chloroform / isoamyl alcohol, and gently invert and mix until the solution becomes milky. Centrifuge at 12000rpm for 2min to separate the phases. Transfer the supernatant to a clean centrifuge tube, add 5 μl ...

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Abstract

A process for separating residual DNA from soy, soybean milk and tomato sass includes centrifugal treating, taking deposit, dissolving in CTAB solution, waving, adding phenol or chloroform, stirring, centrifugal separation, applying supernatant to sephadex-G50 column, adding isopropanol, centrifugal separation, washing deposit with alcohol, centrifugal separation, washing, drying, adding Chelax solution, stirring, boiling, and centrifugal separation. Its supernatant can be directly used as the template of PCR.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for isolating DNA from soy sauce, soybean milk and tomato paste. Background technique [0002] Since the birth of the world's first transgenic plant in 1983, modern biotechnology centered on transgenic technology has developed rapidly. Foods that use genetically modified organisms as direct food or processed as raw materials are genetically modified foods. At present, most genetically modified foods come from genetically modified crops, which have made great contributions to human beings. They can improve food quality, resist pests and diseases, increase production and reduce the use of pesticides, thereby bringing significant economic and social benefits. Today, when returning to nature and advocating organic food, consumers still have doubts about the safety of genetically modified food. The controversy brought about by genetically modified crops and food is u...

Claims

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Application Information

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IPC IPC(8): C07H21/04C12N15/11C12P19/34
Inventor 董海涛李德葆姜玉新
Owner ZHEJIANG UNIV