Side sequence of exogenous insert of transgene paddy strain Kemingdao 1
A technology of transgenic rice and flanking sequence, which is applied in the direction of genetic engineering, plant genetic improvement, determination/inspection of microorganisms, etc.
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Embodiment 1
[0030] Cloning of the flanking sequence of the foreign insert of the transgenic rice line KMD1 (KMD1)
[0031] 1. Experimental materials
[0032] 1. Plant materials
[0033] Transgenic rice: KMD1, a genetically modified rice line.
[0034] Conventional rice: Xiushui 11.
[0035] 2. Enzymes and reagents
[0036] Molecular biology reagents, such as dNTPs, Taq DNA polymerase, and DL2000Marker were purchased from Dalian Bao Biological Biological Engineering Co., Ltd.
[0037] Other biochemical reagents are imported aliquots or domestically produced analytical pure. The primers and degenerate primers were synthesized by Beijing Aoke Biotechnology Co., Ltd.
[0038] 3. Experimental equipment
[0039] PCR thermal cycler: Eppendorf Mastercycle gradient (Eppendorf co.)
[0040] Nucleic acid electrophoresis instrument: DYY-III nucleic acid electrophoresis instrument (Beijing Liuyi Instrument Factory)
[0041] DNA sequencer (Applied Biosystem 377 automatic fluorescence sequence analyzer)
[0...
Embodiment 2
[0082] Qualitative PCR detection method based on the flanking sequence of the foreign insert of the transgenic rice line KMD1 (KMD1)
[0083] 1. Experimental materials
[0084] 1. Plant materials
[0085] Transgenic rice: KMD1, a genetically modified rice line.
[0086] Conventional rice: Xiushui 11.
[0087] 2. Enzymes and reagents
[0088] Molecular biology reagents, such as dNTPs, Taq DNA polymerase, and DL2000Marker were purchased from Dalian Bao Biological Biological Engineering Co., Ltd. Other biochemical reagents are imported aliquots or domestically produced analytical grades. The primers were synthesized by Beijing Aoke Biotechnology Co., Ltd.
[0089] 3. Experimental equipment
[0090] PCR thermal cycler: Eppendorf Mastercycle gradient (Eppendorf co.)
[0091] Nucleic acid electrophoresis instrument: DYY-III nucleic acid electrophoresis instrument (Beijing Liuyi Instrument Factory)
[0092] DNA electrophoresis analysis system: Kodak EDAS 290 (Kodak co.)
[0093] Other in...
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