Standard plasmid molecular used for detecting transgenic soybean, corn and cotton and construction of the standard plasmid molecular
A technology of transgenic soybean and transgenic corn, which is applied in the field of plasmid molecules in the field of molecular biology technology, can solve the problem of difficulty in obtaining positive standard samples of transgenic crops, and achieve the effects of high sensitivity, high specificity and repeatability
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0028] Example 1: Construction of standard plasmid molecules
[0029] 1. Experimental materials
[0030] Transgenic maize lines Bt176 and Mon810; non-transgenic maize varieties.
[0031] 2. Experimental reagents
[0032] pMD TM 19-T Simple Vector was purchased from Dalian Bao Bioengineering Company; dNTPs, Taq DNA polymerase, DNA marker I, II and marker III were purchased from Beijing Quanshijin Biotechnology Co., Ltd.; restriction endonucleases Not I, Sal I, EcoR I and Xba I were purchased from NEB Beijing Co., Ltd.; plasmid genomic DNA was extracted and purified using a plasmid mini-extraction kit developed by Beijing Tiangen Biochemical Technology Co., Ltd.; other biochemical reagents were imported subpackages or domestic analytical grades.
[0033] 3. Experimental equipment
[0034]TC-512 PCR Amplifier (TECHNE, UK); Geliance 200 DNA Electrophoresis Gel Imager (PerkinElmer, USA); Nano-Drop ND-1000 Nucleic Acid Protein Analyzer (Nano Drop, USA); centrifuge; constant tem...
Embodiment 2
[0066] Example 2: Application of constructed standard plasmid molecules in actual detection
[0067] 1. Enzymes and Reagents
[0068] Real-time PCR detection kit (Premix Ex Taq TM (Perfect Real Time)) were purchased from Bao Bioengineering (Dalian) Co., Ltd.; primers and TaqMan probes were synthesized by Shanghai Jikang Biotechnology Co., Ltd.; other biochemical reagents were imported subpackage or domestic analytical grade.
[0069] 2. Experimental equipment
[0070] Real-time fluorescence quantitative PCR instrument 7500 (ABI company).
[0071] 3. Experimental method and process
[0072] 1. Design and synthesis of fluorescent quantitative PCR primers and probes
[0073] Primers and probes for quantitative PCR were designed using the software Primer Premier 5.0 according to the transformation event sequences at the 3' ends of Hmg and Bt176 and Mon810 strains in standard plasmid molecules, as shown in Table 4. The 5' end of the TaqMan probe is labeled with the reporter fl...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com