Preparation method of Magnaporthe oryzae mycelium holoprotein two-dimensional electrophoresis sample
A rice blast fungus, two-dimensional electrophoresis technology, applied in the biological field to achieve the effect of prolonging the treatment time
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Embodiment 1
[0029] 1) Transfer the blast fungus strain from the test tube to the solid medium, culture in dark at 28°C for 7 days, and when the diameter of the colony reaches 4-6cm, cut out 3-5 pieces of mycelium with a size of 0.5cm×0.5cm, and transfer to Put it on 50mL rice blast fungus full nutrient liquid medium, 150r.min at 28°C -1 The shaker was shaken for 3 days, and the hyphae were filtered through double-layer gauze, and then washed with sterile water to remove the culture medium.
[0030] 2) Weigh 5 g of wet mycelia and transfer to 50 ml of freshly prepared full-nutrient liquid medium. 150r.min at 28°C -1 Shake the shaker for 48 hours, filter the culture with gauze, wash the mycelium with sterile water to remove the medium after filtering through double-layer gauze, drain the clean mycelium with a suction filter bottle, and place the dried mycelium in Store in -80°C refrigerator for later use.
[0031]3) Weigh 1 g of mycelium, grind it in liquid nitrogen with a mortar and sus...
Embodiment 2
[0044] 1) Transfer the blast fungus strain from the test tube to the solid medium, culture in dark at 28°C for 7 days, and when the diameter of the colony reaches 4-6cm, cut out 3-5 pieces of mycelium with a size of 0.5cm×0.5cm, and transfer to Put it on 50mL rice blast fungus full nutrient liquid culture medium, shake it at 28°C for 3 days at 150r.min-1, filter the hyphae through double-layer gauze, and wash with sterile water to remove the culture medium.
[0045] 2) Weigh 5 g of wet mycelia and transfer to 50 ml of newly prepared nitrogen-starved liquid medium. 150r.min at 28°C -1 Shake the shaker for 48 hours, filter the culture with gauze, wash the mycelium with sterile water to remove the medium after filtering through double-layer gauze, drain the clean mycelium with a suction filter bottle, and place the dried mycelium in Store in -80°C refrigerator for later use.
[0046] 3) Weigh 1 g of mycelium, grind it in liquid nitrogen with a mortar and suspend it in a certain...
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