Plasmid reference molecule of quantitative determination of nucleic acids from transgenic soybean GTS40-3-2
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A technology of genetically modified soybeans and standard molecules, applied in the field of bioengineering, can solve the problem of lack of positive standard product configuration for positive products, and achieve the effect of good traceability
Inactive Publication Date: 2012-06-27
SHANGHAI INST OF MEASUREMENT & TESTING TECH
View PDF4 Cites 9 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
[0008] The technical problem to be solved by the present invention is to provide a plasmid standard for transgenic soybean GTS40-3-2 in order to overcome the lack of positive products and the configuration of positive standard products in the existing nucleic acid quantitative PCR detection of transgenic soybean GTS40-3-2 molecular
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more
Image
Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0073] The construction of embodiment 1 plasmid standard molecule
[0074] 1. Experimental reagents
[0075] Restriction endonucleases KpnI, HindIII, XbaI, and restriction enzyme buffers were purchased from Shanghai Haojia Technology Development Co., Ltd.
[0076] pUC19 vector, Taq DNA polymerase, T4 DNA ligase, dNTP, DL2000 Marker
[0077] Purchased from Shanghai Haojia Technology Development Co., Ltd.;
[0078] DNA primers were synthesized by Shanghai Sangon Biotechnology Co., Ltd.
[0079] Other biochemical reagents are imported subpackages or domestic analytical pure.
[0083] DNA electrophoresis analysis system, including dark box, digital camera, computer, scanner, inkjet printer, photosensitive printer, Tanon UV-2000 ultraviolet analyzer, Gis gel image analysis software (Sha...
Embodiment 3
[0121] Application of the plasmid standard molecule pXL02 constructed in embodiment 3 in actual detection
[0122] 1. Experimental reagents
[0123] Plant genomic DNA was extracted and purified using the Plant Genomic DNA Extraction Kit from OMEGA.
[0124] Plasmid DNA was extracted and purified using a plasmid DNA extraction kit from OMEGA.
[0125] The pUC19 vector, T4 DNA ligase and its buffer, dNTPs, Taq DNA polymerase and its buffer, and DNA Marker were purchased from Shanghai Haojia Technology Development Co., Ltd.
[0126] Primers and probes were synthesized by Shanghai Sangon Bioengineering Co., Ltd.
[0127] Other biochemical reagents are imported subpackages or domestic analytical pure.
[0130] Opticon-2 Quantitative PCR Amplifier (MJ Research Inc.).
[0131] DNA electrophoresis analysis system, including dark box, dig...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more
PUM
Login to view more
Abstract
The invention discloses a plasmid reference molecule which is suitable for the quantitative determination of nucleic acids from a transgenic soybean GTS40-3-2, comprising a structure-specific sequence and a strain-specific sequence of the transgenic soybean GTS40-3-2 and a specific fragment of a soybean endogenous reference gene Lectin. The invention is characterized by designing primers to obtain the structure-specific sequence and the strain-specific sequence of the transgenic soybean GTS40-3-2 and the specific fragment of the soybean endogenous reference gene Lectin by PCR amplification through the analysis of inserting the exogenous source of the transgenic soybean GTS40-3-2 in the gene sequence; constructing by molecular cloning to form a artificial recombinant plasmid molecule pXL02 in a plasmid molecule; detecting the phosphorus content of the plasmid reference molecule by inductively coupled plasma mass spectrometry (ICP-MS) to calculate the concentration of the plasmid reference molecule. According to the invention, the plasmid reference molecule constructed in the invention can completely replace a transgenic soybean GTS40-3-2 positive standard sample, and the plasmid reference molecule is completely suitable for the quantitative PCR analysis and detection of structure specificity and strain specificity of the transgenic soybean GTS40-3-2 sample.
Description
technical field [0001] The invention relates to a plasmid molecule in the technical field of bioengineering, in particular to a plasmid standard molecule for quantitative detection of genetically modified soybean GTS40-3-2 and its construction and quantitative method. Background technique [0002] Genetically modified biotechnology is known as "the most rapidly applied biotechnology". With the continuous development of biotechnology, more and more genetically modified plants are used in the agricultural field to promote agricultural production, help farmers increase income, and solve the problems caused by global population growth. The food problem brought about opened up new avenues. The so-called transgenic plant (Genetically modified organism, GMO) refers to the use of biotechnology to recombine genes isolated or artificially synthesized from different organisms in vitro, and then introduce them into recipient plant cells, so that the new genes can be integrated in the re...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more
Application Information
Patent Timeline
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to view more 
Login to view more