Transgene agricultural product DNA detection chip, its preparation method and application

A technology for detecting chips and agricultural products, applied in biochemical equipment and methods, microbe measurement/inspection, etc., can solve the problem that chips cannot get rid of cost

Inactive Publication Date: 2008-02-20
中国疾病预防控制中心营养与食品安全所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Transgene agricultural product DNA detection chip, its preparation method and application
  • Transgene agricultural product DNA detection chip, its preparation method and application
  • Transgene agricultural product DNA detection chip, its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1, Preparation of Transgenic Agricultural Products Detection Chip

[0064] 1. Extraction of genomic DNA of transgenic agricultural products (CTAB method and silica gel column purification method)

[0065] Samples and sources of genetically modified agricultural products: positive and negative samples of genetically modified soybean 40-3-2, genetically modified corn MON810, GA21, and NK603 were provided by Monsanto. Positive and negative samples of transgenic rapeseed Ms1Rf1 and T45 were provided by Bayer Company. S86-positive and negative samples of transgenic rice SCK were provided by the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences.

[0066] Cetyltrimethylammonium nitrate (CTAB) extraction buffer preparation method: 20g / l CTAB, 4.0g; 4M NaCl, 16.4g; 0.1M Tris-HCl, 3.15g; 20mM sodium edetate (Na 2 EDTA), 1.5g; add 100ml of sterile deionized water, adjust the pH to 8.0 with 1M NaOH, and finally adjust the volume to 200ml and au...

Embodiment 2

[0117] Embodiment 2, the preparation of test sample

[0118] 1. Genomic DNA extraction method of the sample to be tested

[0119] For the method, refer to the "Method for Extracting Genomic DNA from Transgenic Agricultural Products" in Example 1.

[0120] 2. Labeling of samples to be tested (multiple PCR fluorescein labeling)

[0121] 25 μ l volume: PCR buffer solution, 2.5 μ l; Primer (mixed primer) table 2 and 3 listed mixed primer concentration and mixing ratio see the following specific examples); dNTP (10mM, no dTTP), 0.5 μ l; Cy5-dUTP ( 1mM) (Pharmacia), 0.5μl; dTTP (1mM), 2.0μl; template DNA (genomic DNA of the sample to be tested above), 1.0-2.0μl; Taq (5U / μl), 0.3-0.5μl; Make up to 25μl.

[0122] PCR reaction conditions: with embodiment 1.

Embodiment 3

[0123] Embodiment 3, the detection of product to be tested

[0124] 1. Hybridization and washing of slides

[0125] Steps:

[0126] (1) Place the slide in the hybridization chamber.

[0127] (2) Heat and denature the microcentrifuge tube containing the labeled probe: cook in boiling water at 100° C. for 3 minutes.

[0128] (3) Add 100 μl of high-efficiency hybridization solution (Dingguo Company) into the tube, mix well by pipetting and pour into the hybridization chamber. Hybridize at 42°C for 12-18 hours.

[0129] (4) After 12-18 hours, the slides were taken out, and put into membrane washing solution I (2×SSC, 0.1% SDS) to wash for 2 minutes.

[0130] (5) Wash the membrane with II solution (0.1×SSC, 0.1% SDS) for 2 minutes.

[0131] (6) Wash with sterile high-purity water for 2 minutes, dry and wait for scanning.

[0132] 2. Observe the hybridization results under the scanner

[0133] The test results were judged based on the signals read by a scanner (Perkin Elmer, ...

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Abstract

The invention relates to a transgene farm product DNA detecting chip. The chip includes film base and nucleic acid probe. The probe contains target detection probe. The target detection probe is a series of nucleotide sequence showed as SEQ ID NOs.1-50. The invention offers the making method, application, kit and detecting method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a DNA detection chip for genetically modified agricultural products and its preparation method and application. Background technique [0002] The use of modern biotechnology to cultivate genetically modified crops (GMO for short) that are different from traditional breeding has created a new era for agricultural research and opened an unprecedented opportunity for humans to improve the quantity and quality of food. . Modern biotechnology, also known as recombinant DNA technology, has great potential in improving the nutritional quality of food, increasing crop yield and disease resistance, insect resistance, reducing the amount of pesticides, and vaccination against human infectious diseases. In recent years, genetically modified crops have become a hot spot for research and development in various countries. In recent years, the global planting area and sales of transgenic plants h...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 吴永宁张建中周萍萍
Owner 中国疾病预防控制中心营养与食品安全所
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