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DNA chip suitable for high-throughput detection of transgenic products

A DNA chip and genetically modified technology, applied in the field of DNA chips, can solve problems such as methods that need to be further improved, expensive liquid phase chip instruments, and prone to false positive results, so as to improve detection accuracy, reduce detection costs, and shorten detection time. Effect

Active Publication Date: 2014-03-26
SHANGHAI JIAO TONG UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

The combination of the two is a new trend in GMO detection, but many experiments have found that false positive results are prone to occur when there are more than 10 samples to be tested (Chaouachi M, et al, 2008, Agric.Food Chem., 56, 11596-11606), The method needs to be further improved
In theory, microspheres can be combined with any primer / probe combination to better meet the needs of high-throughput detection, but the disadvantage of liquid-phase chips is that it is still in further research, and expensive liquid-phase chip instruments need to be purchased

Method used

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  • DNA chip suitable for high-throughput detection of transgenic products
  • DNA chip suitable for high-throughput detection of transgenic products
  • DNA chip suitable for high-throughput detection of transgenic products

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Designing appropriate primers and probes is the key to accurate GMO detection. Design specific primers and probes for the 97 target sequences (41 common insertion elements, 48 ​​common events and 8 endogenous genes) mainly contained in the current transgenic products, and the designed primers are verified by conventional PCR for specificity , multiple probes designed were screened with genome templates, and the most accurate primers and probes were finally determined.

[0045] The seed powder of 4 kinds of transgenic soybean events (A2704-12, A5547-127, GTS40-3-2 and MON89788) in the laboratory was extracted and purified using a commercial DNA extraction kit, and then the NanoDrop1000 The concentration of DNA was measured and uniformly diluted to 10ng / ul, and 10ul of each of the four templates was fully mixed as the sample to be tested.

[0046] Chip PCR reaction was carried out by using hydrophilic and hydrophobic microporous chip technology. That is, the designed 97...

Embodiment 2

[0050] The mixed samples of 8 kinds of transgenic rice (TT51-1, Kefeng NO.6, Kefeng NO.8, T2A-1, KMD1, Bar68-1, t1c-19 and RJ5) were detected by the above experimental process, and the chip scanning results are shown in the attached image 3 ;Depend on image 3 and combine the pattern figure 1 It can be seen that the probe signals in the hybridization graph are: 1, 2, 3, 6, 8, 15, 19, 20, 21, 24, 26, 28, 29, 33, 39, 81, 82, 83, 84 , 85, 86, 87, 88 and 95, the corresponding target names are: P-CaMV35s, bar, bla, T-nos, Pat, nptII, Pact, P-Ubi, POrbcs, cry1Ab / c, CpTI, cry1C, CMoVb, uidA, Gus, TT51-1, Kefeng NO.6, Kefeng NO.8, T2A-1, KMD1, Bar68-1, t1c-19, RJ5 and rice endogenous gene Sps. According to the known 8 kinds of transgenic soybean events, and searching relevant databases and literatures, it is concluded that the theoretical 8 events and rice endogenous signals are completely detected, and the component signals are only 8 and 29 more than the theoretical signals. The...

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Abstract

The invention discloses a DNA chip suitable for high-throughput detection of transgenic products. A specific DNA probe capable of detecting 97 transgenic targets is fixed on a vector of the DNA chip, and the nucleic acid sequences of the DNA probe are as shown in SEQ ID NO.1 to SEQ ID NO.97. The 97 oligonucleotide probes of the DNA chip disclosed by the invention are designed for the most widespread transgenic insertion sequences and endogenous genes of 8 crop varieties and have high specificity; high-density DNA chip technology can be used for simultaneously detecting the target sequences of a plurality of transgenic components in a sample, and the DNA chip has the advantages of high efficiency, high accuracy and high sensitivity. The DNA chip disclosed by the invention has a wide application range in the field of transgenic product detection and identification management.

Description

technical field [0001] The invention relates to a detection method in the technical field of bioengineering, in particular to a DNA chip suitable for high-throughput detection of transgenic products. Background technique [0002] GMOs (Genetically Modified Organisms, GMOs) refer to a class of biological organisms whose genetic material is not changed through mutual hybridization or natural integration under natural conditions. A GMO must itself be a biological entity that can reproduce or transmit genetic material. In a broad sense, GMO includes all life forms of genetically modified organisms. In a narrow sense, it refers to genetically modified plants and animals. Since genetically modified animals have not been commercialized except for ornamental fish and pets, they strictly refer to genetically modified plants (Li Quanfen et al., 2011, Chinese Animal Husbandry and Veterinary Medicine, 38, 152). [0003] Since 1994, when the first genetically modified tomato FLAVR SAVR...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C40B40/06
CPCC12Q1/6895C12Q2600/13C12Q2600/156C12Q1/6837
Inventor 杨立桃陶生策张大兵邵宁蒋诗萌
Owner SHANGHAI JIAO TONG UNIV
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