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DNA extracting method and use thereof in detecting transgene component in food

A technology of genetically modified ingredients and samples, applied in the field of DNA extraction, can solve problems such as the quality cannot meet the requirements, and achieve the effect of high quality

Inactive Publication Date: 2009-04-22
HAI KANG LIFE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The quality of DNA extracted by this method still cannot meet the requirements, and the obtained DNA product still contains many impurities that inhibit PCR.

Method used

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  • DNA extracting method and use thereof in detecting transgene component in food
  • DNA extracting method and use thereof in detecting transgene component in food
  • DNA extracting method and use thereof in detecting transgene component in food

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1. Amplification and detection of cauliflower mosaic virus 35S promoter sequence in transgenic food

[0053] Materials and methods

[0054] 1. Samples: Soybeans (USA), soybeans (Mexico), cup noodles 1, cup noodles 2, bag noodles 1, bag noodles 2 and soybean milk (Hong Kong) used in the present invention are all commercially available products.

[0055] 2. Reagents:

[0056] DNA extraction reagents:

[0057] Buffer A: 10 mM Tris HCl pH 8, 150 mM NaCl, 2 mM EDTA. 1% SDS.

[0058] Buffer B: 5M Guanidine Hydrochloride

[0059] PCR Super Mix

[0060] Element Stock solution concentration Final concentration PCR buffer 10× 1× MgCl 2 50mM 2.1mM DNTPs 10mM 0.2mM

[0061] Wherein, the PCR buffer is a commonly used Tris-HCl buffer.

[0062] Taq polymerase: purchased from Invitrogen USA.

[0063] Primers: purchased from Invitrogen USA, the sequences of which are shown in SEQ ID No.1 and 2 in the sequence listing.

[0064] ...

Embodiment 2

[0076] Example 2. Amplification and detection of Agrobacterium tumefaciens nopaline synthase (NOS) terminator

[0077] The DNA extraction method and PCR amplification and detection method are the same as in Example 1. The samples are 1. Seasoning powder 1, 2. Seasoning powder 2, 3. Seasoning powder 3, 4. Seasoning powder 4, 5. Bowl of noodles 1, 6. Bowl of noodles 2, 7. Instant macaroni noodles, 8 . Instant Macaroni. The primers used therein are shown in SEQ ID Nos.6 and 8. The positive control was changed to IRMM410 * . The negative control was DEPC water. The result is as Figure 7 shown. The result is that samples 5. Bowl of noodles 1, 6. Bowl of noodles 2, 7. Instant macaroni noodles, and 8. Instant macaroni noodles have positive 146bp bands. It shows that there are genetically modified ingredients in these foods.

Embodiment 3

[0078] Example 3. Amplification and detection of neomycin phosphotransferase II (NPTII) sequence

[0079] The DNA extraction method and PCR amplification and detection method are the same as in Example 1. The samples used are 1. rapeseed oil 1, 2. rapeseed oil 2, 3. rapeseed oil 3, 4. rapeseed oil 4, 5. rapeseed oil 5. The primers used therein are shown in SEQ ID Nos.11 and 14, and the positive control is IRMM410. The result is as Figure 8 shown. The result was a positive band of 194bp in samples 3 and 4, indicating that there were genetically modified ingredients in these two foods.

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Abstract

A process for extracting DNA used to detecting the transgenic components in food by PCR method includes treating sample by cracking liquid, centrifugal separation to obtain supernatant, filtering, adding extractant, centrifugal separation to obtain supernatant, adding resin, adsorbing DNA, eluting, centrifugal separation to obtain filtrate containing DNA, and precipitating DNA.

Description

technical field [0001] The invention relates to a DNA extraction method and its application in the detection of genetically modified food. Background technique [0002] Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) are contained in food, and people ignored their existence before. Until recent years, many GM crops were approved for cultivation in Europe and elsewhere. Nucleic acid has now become an important tool for analyzing food in food analysis. In several recent EU laws and regulations, the sale of food containing genetically modified ingredients must be labeled. Many countries have adopted or are considering introducing laws to promote the labeling of foods containing genetically modified ingredients. [0003] Therefore, it is necessary to develop appropriate techniques for identifying which foods must be labeled. In practice, the best way to distinguish GMOs from non-GMOs is to prove the presence or absence of foreign genes at the DNA level. Polymerase ch...

Claims

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Application Information

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IPC IPC(8): C12N15/10C07H21/04C12Q1/68
Inventor 于常海刘乐庭高龙生
Owner HAI KANG LIFE
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