New lycine aldehyde dehydrogenase gene BnBADH-1 in cabbage type rape
A technology of betaine aldehyde dehydrogenase and Brassica napus, which is applied in the directions of genetic engineering, plant genetic improvement, oxidoreductase and the like
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[0009] The present invention will be described in detail below in conjunction with specific embodiments.
[0010] ① Obtaining sterile seedlings: Soak Brassica napus seeds in 75% ethanol for 1 min and 0.1% mercuric chloride for 8 min to sterilize them, wash them repeatedly with sterilized water for 3 to 5 times, and then place them on MS solid medium; 25°C in an artificial climate chamber, 70% relative humidity, and 16h light / 8h dark conditions for growth.
[0011] ② Extraction of rapeseed total RNA and synthesis of total cDNA: Take rapeseed leaves about 3 weeks after germination, use Trizol kit to extract total RNA; use RNase-free DNase I to remove residual DNA from the obtained total RNA, and then reverse The first-strand cDNA was generated using the transcription kit: Oligo(dT)18 primer was used for reverse transcription, and after incubation at 42 °C for 30 min, the enzyme was inactivated by heating at 85 °C for 5 min. The resulting cDNA was stored at -20°C for subsequent ...
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