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Biology: Arabidopsis Culture Cell and Transformation

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Arabidopsis Culture Cell and Transformation Arabidopsis thaliana (L.) Columbia ecotype suspension- cultured T87 cells were maintained at 22°C in JPL3 medium with continuous illumination and shaking at 100g. Two-week-old cells were sieved through 500 μm stainless mesh and the remaining filtrate was transferred to a flask containing 20 ml of fresh JPL3 medium for subculture. Transformation of T87 cells was done by culturing the cells in B5 medium supplemented with 1 μM 1-naphthaleneacetic acid (NAA) and 40 g L-1 sucrose. The cells were cultured for one day at 22°C with continuous illumination and shaking at 120g. Next, 10 μL of overnight cultured Agrobacterium transformed with respective vectors were added into the cell suspension and cultured for an additional two days. After co-cultivation, the cell suspension was washed thrice with 10 mL of JPL3 medium supplemented with Carbencilin (250 μg mL-1) by centrifuging at 100g for two minutes. Finally, the cells were resuspended and spread onto JPL3 selection agar plate supplemented with Carbencilin (250 μg mL-1), Kanamycin (30 μg mL-1) an...
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