Method for screening transgenic rape using chanamyn seed soaking method
A technology of kanamycin and transgene, which is applied in the field of screening transgenic rapeseed by kanamycin seed soaking method, can solve the problems of heavy workload, complicated operation, false positive, etc., and achieve fast screening speed, less false positive, and simple method Effect
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Embodiment 1
[0017] The screening of transgenic seeds of Chinese cabbage-type rapeseed, the specific screening process is as follows:
[0018] (1) Preparation of kanamycin solution Dissolve 200mg of kanamycin solid in 1L of water to obtain a 0.2% kanamycin solution;
[0019] (2) Seed soaking 10,000 T. 1 Substitute seeds (from the Rapeseed Research Center of Sichuan Agricultural University) were put into kanamycin solution and soaked for 16 hours, and were taken out when they turned white;
[0020] (3) Screening Seeds are sown on soil that does not contain kanamycin, soak the soil before sowing, and directly sow on the soil surface, observe the results after 2-3 days, select normal green seedlings, and get 12 transgenic plants .
Embodiment 2
[0022] The screening of Brassica napus transgenic plants, the specific screening process is as follows:
[0023] (1) prepare kanamycin solution, 280mg kanamycin solid is dissolved in 1L water, get concentration and be the kanamycin solution of 0.28%;
[0024] (2) Seed soaking: 20,000 grains of transgenic T seeds marked by the kanamycin resistance gene obtained by the floral-dip method (translated as inflorescence soaking method) 1 Substitute seeds (from the Rapeseed Research Center of Sichuan Agricultural University) were soaked in a kanamycin solution for 16 hours, the solution just submerged the seeds, and were taken out when they turned white;
[0025] (2) Screening Soak the soil before sowing, and sow the seeds on the soil without kanamycin; observe the results after 2-3 days, select normal green seedlings, and obtain 20 transgenic plants.
Embodiment 3
[0027] Screening of Transgenic Plants of Brassica napus Using Traditional Medium Containing Kanamycin
[0028] Proceed as follows:
[0029] (1) The screened materials are transgenic seeds of Brassica napus (from the Rapeseed Research Center of Sichuan Agricultural University).
[0030] (2) Preparation of culture medium Prepare MS medium containing 270 mg / L kanamycin according to the usual method, and mark 1500 grains obtained by the floral-dip method (translated as inflorescence soaking method) with the kanamycin resistance gene transgenic T 1 The generation seeds were sterilized and inoculated into the medium for screening, and the results were observed one week later.
[0031] (3) Screening Normal green seedlings were selected to obtain 18 transgenic plants.
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