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Probe combination, liquid-phase chip and kit for transgenic corn detection, and application thereof

A technology for the detection of genetically modified corn and probes, which is applied in the direction of recombinant DNA technology, measurement/inspection of microorganisms, DNA/RNA fragments, etc., which can solve the problems of increased difficulty in detection and fewer types of genetically modified ingredients, and achieve high sensitivity and high detection results. Reliable, the effect of many indicators that can be detected in a single time

Active Publication Date: 2017-03-15
上海泽衡生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the large variety and quantity of genetically modified crops, many of them are imported as processing raw materials. After treatment, the genetically modified ingredients may be degraded, so the detection becomes more difficult, and the existing detection kits can detect the types of genetically modified ingredients in a single test few

Method used

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  • Probe combination, liquid-phase chip and kit for transgenic corn detection, and application thereof
  • Probe combination, liquid-phase chip and kit for transgenic corn detection, and application thereof
  • Probe combination, liquid-phase chip and kit for transgenic corn detection, and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0044] The probe combination provided in this example for the detection of transgenic maize includes a 5' terminal labeled C12 molecular arm and an amino-modified positive probe and a detection probe. The base sequence of the positive probe is shown in SEQ ID NO.33, and the base sequences of the detection probes are respectively shown in SEQ ID NO.23-SEQ ID NO.32.

[0045] Wherein, the gene component detected by the positive probe is maize invertase I gene (Irv1), which is an internal reference gene and serves as a positive control.

[0046] The 5' end of the detection probe is also labeled with C12 molecular arms and amino modifications for conjugation to fluorescently encoded microspheres. There are 10 detection probes in total, which are the 1st to 10th detection probes, the base sequences of which are shown in SEQ ID NO.23-SEQ ID NO.32.

[0047] Wherein, the first detection probe whose base sequence is shown as SEQ ID NO.23 is used to detect the glufosinate acetyl-CoA tra...

Embodiment 2

[0062] The probe combination provided by the present invention for the detection of transgenic maize is the same as in Example 1, but the different probe combinations of the present invention also include a 5' end marker C 12 Molecular arm and amino-modified negative probe, the base sequence of the negative probe is shown in SEQID NO.34. The probe combination provided by the present invention for the detection of transgenic maize not only has the same effect as in Example 1. There are other effects reflected in that the negative probe shown in SEQ ID NO.34 does not combine with any PCR amplified product during the detection process, which can play a good negative control effect and further avoid false positive results. occur, improving the reliability of the test results.

Embodiment 3

[0064] The liquid phase chip for the detection of transgenic corn provided by this embodiment, the liquid phase chip is composed of the probe combination (i.e. the positive probe and the 1-10th detection probe) for the detection of genetically modified corn described in Example 1 respectively Conjugated with fluorescently encoded microspheres. The coupling steps are described in detail below. However, the liquid phase chip of the present invention is not limited to the coupling method described below, and can also be produced by other coupling methods, as long as any of the probe combinations described in Example 1 is used Combining with fluorescent coded microspheres to realize liquid phase chip detection of corn transgenic components falls within the scope of protection of the present invention. This example can be prepared by the following coupling method.

[0065] 1 Prepare the coupling system: take 40 μl (the total number of microspheres is 1×10 5 a) hydroxylated fluor...

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Abstract

The invention provides probe combination, a liquid-phase chip and a kit for transgenic corn detection, and application thereof, and relates to the technical field of transgenic detection. The probe combination comprises 10 detection probes and a positive probe, wherein the 10 detection probes are designed according to common exogenous genes Pat, Bar, Cry1Ab, Cry1Ac, Cry105, EPSPS-P0, EPSPS-P2, P35S, T35S and Nos-3' in the transgenic corn; and the positive probe is designed according to an endogenous gene Ivr1 of the corn. The probe combination can be used for detecting whether the transgenic corn contain the above transgenic components or not after being coupled with fluorescent encoding microspheres separately, has the characteristics of simplicity in operation, high specificity, high sensitivity and large flux, and has an important function on transgenic corn detection of seed stations, institutes of agricultural sciences and border ports.

Description

technical field [0001] The invention relates to the technical field of transgenic detection, in particular to a probe combination, a liquid phase chip, a kit and applications thereof for the detection of transgenic corn. Background technique [0002] At present, genetically modified corn is one of the most important genetically modified food crops in the world. According to the International Service for the Application of Agricultural Biotechnology (ISAAA), the cumulative planting area of ​​genetically modified crops in the world during the 20-year period from 1996 to 2015 reached an unprecedented 2 billion hectares, which is equivalent to the total area of ​​China (956 million hectares) or the United States. Twice the total area (937 million hectares), of which GM corn accounted for 30%. A large number of genetically modified crops appear in the global market. China, the European Union and other countries and organizations require their labeling when their environmental an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 郭尚秦子苏王岳华周均
Owner 上海泽衡生物科技有限公司
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