Exogenous insertion element flanking sequences of transgenic rice strain 223F-S21 and application thereof
A technology of 223F-S21 and transgenic rice, which is applied in the direction of DNA/RNA fragments, microbe determination/inspection, DNA preparation, etc., and can solve the problem of side sequence of foreign insert fragment without transgenic rice line 223F-S21, etc.
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Embodiment 1
[0058] Example 1, Cloning of the flanking sequence of the exogenous insertion vector of the transgenic rice line 223F-S21
[0059] 1. Experimental materials
[0060] 1. Plant material
[0061] Transgenic rice: transgenic rice line 223F-S21.
[0062] 2. Enzymes and reagents
[0063] Molecular biology reagents, such as Taq enzyme, TaKaRaLA Taq, pMD18-T, and DNA Marker were purchased from Dalian Bao Biological Engineering Co., Ltd.
[0064] Other biochemical reagents are imported or domestic analytically pure. Primers and degenerate primers were synthesized by Shanghai Sangong.
[0065] 3. Experimental equipment
[0066] PCR amplification (Eppendorf co.), nucleic acid electrophoresis instrument (DYY-6C, Beijing Liuyi Instrument Factory)
[0067] Other instruments include: centrifuge, electric heating constant temperature water tank, electronic balance, incubator, etc.
[0068] 2. Experimental method and process
[0069] 1. Rice genomic DNA extraction and detection
[007...
Embodiment 2
[0109] Qualitative PCR detection of the flanking sequence of the foreign insert in transgenic rice 223F-S21
[0110] 1. Experimental materials
[0111] 1. Plant material
[0112] Transgenic rice: Transgenic rice line 223F-S21
[0113] Conventional rice: Xiushui 134
[0114] 2. Enzymes and reagents
[0115] Molecular biology reagents, such as Taq enzyme, TaKaRaLA Taq, pMD18-T, and DNA Marker were purchased from Dalian Bao Biological Engineering Co., Ltd.
[0116] Other biochemical reagents are imported or domestic analytically pure. Primers and degenerate primers were synthesized by Shanghai Sangong.
[0117] 3. Experimental equipment
[0118] PCR amplification (Eppendorf co.), nucleic acid electrophoresis instrument (DYY-6C, Beijing Liuyi Instrument Factory)
[0119] Other instruments include: centrifuge, electric heating constant temperature water tank, electronic balance, incubator, etc.
[0120] 2. Experimental method and process
[0121] 1. Rice genomic DNA extrac...
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