Simple, efficient and real-time transgenic product detecting method and application thereof
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A detection method and genetically modified technology, applied in the biological field, can solve problems such as complex costs
Inactive Publication Date: 2018-03-30
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However, the current DNA-based PCR detection methods all require PCR amplification, which is complicated and costly; therefore, it is necessary to develop a simple, real-time, low-cost transgenic method that can be realized by non-professionals in a "fool" mode. Product testing method
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Embodiment 1
[0038] This embodiment provides a simple, efficient and real-time detection method for genetically modified products, including the following steps.
[0039] Tissue processing: Take appropriate samples of product A and product B respectively, and quickly grind to homogenize.
[0040] Nucleic acid extraction: The magnetic bead method is used for nucleic acid extraction, and the specific combination of magnetic beads and nucleic acid is used to extract high-purity and high-concentration nucleic acid.
[0041] Target DNA capture: Denature the extracted nucleic acids, and then use specificity to design and extract chips for transgene capture and extraction based on the principle of Q gene and probe hybridization matching for the target exogenous gene Q gene commonly transferred in the transgenic field. Preparation, and then use the transgene capture extraction chip to capture, extract and enrich the target gene.
[0042] Wherein, the Q gene sequence is the base code sequence repr...
Embodiment 2
[0054] This embodiment provides a simple, efficient and real-time detection method for transgenic products described in the present invention to realize the detection of transgenic insect-resistant rice.
[0055] Tissue processing: Take Huahui No. 1 rice or the Huahui No. 1 rice containing the insect-resistant rice to be tested, and quickly grind to homogenize.
[0056] Nucleic acid extraction: The magnetic bead method is used for nucleic acid extraction, and the specific combination of magnetic beads and nucleic acid is used to extract high-purity and high-concentration nucleic acid.
[0057] Target DNA capture: denature the extracted nucleic acids, and then use the specificity to design and prepare transgene capture extraction chips based on the principle of Bt fusion insecticidal protein gene Cry1Ac / Cry1Ab and probe hybridization matching, and then use transgene capture The extraction chip captures, extracts and enriches target genes.
[0058] Single-molecule sequencing: T...
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Abstract
The invention relates to a simple, efficient and real-time transgenic product detecting method and application thereof. The transgenic product detecting method sequentially comprises the steps that tissue processing is performed, wherein an appropriate number of samples of tissues of agricultural and pastoral plants or animals is taken, ground and homogenized; nucleic acid extraction is performed,wherein a paramagnetic particle method is adopted to perform nucleic acid extraction; targets DNA are captured, denaturating treatment is conducted on extracted nucleic acid, then specificity is utilized to design a transgene capture and extraction chip for a general component for target exogenous genes and / or transgenes generally transformed in the transgene field, and then target genes are captured, extracted and enriched; single molecule sequencing is performed, wherein genetic information of the detected samples is obtained by using a nanopore single molecule sequencing method; result analysis is performed. The simple, efficient and real-time transgenic product detecting method can simply and efficiently achieve detection of transgenic products in real time, the whole process is completed in a full-automatic 'simpleton' way, and accordingly non-professional common practitioners / consumers can also achieve detection of transgenic products.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a simple, efficient and real-time detection method for genetically modified products and its application. Background technique [0002] Genetically modified products are products made of animals, plants or microorganisms with specific foreign genes or gene fragments inserted into cell DNA through non-reproductive methods to obtain certain good traits. In recent years, genetically modified products have increasingly become a hot spot of social concern, so the detection and identification of genetically modified products is becoming more and more important to protect consumers' right to know and right to choose. [0003] The detection methods of genetically modified ingredients can be divided into two categories: qualitative detection and quantitative detection. Qualitative detection methods can detect whether a sample contains genetically modified ingredients and what kind of genetic...
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