Construction method for cornua cervi pantotrichum bioreactor
A technology of bioreactor and construction method, which is applied in biochemical equipment and methods, botanical equipment and methods, cells modified by introducing foreign genetic material, etc., can solve the problems of breast reactor development period limitation, etc. , the effect of increasing production
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Embodiment 1
[0044] 1. Preparation and purification of lentiviral particles
[0045] Take out a certain number of 10cm cell culture dishes, add 4mL of 0.01% poly-lysine to each plate, invert left and right to make the liquid spread on the bottom of the plate, incubate at room temperature for 10min to remove poly-lysine, add 13mL of DMEM to each dish Culture medium (containing 10% FBS) and a certain number of 293t cells were cultured in a cell incubator (37°C, 5% CO 2 ). For each plate of cells, dilute 24 μg DNA with 1.5 mL of serum-free Opti-MEM (pLVTHM:pCMV-dr8.91:pMD2.G plasmid mass ratio is 2:1.5:0.75) and 51 μL with 1.5 mL of serum-free Opti-MEM Transfection reagent Lipofectamine2000, incubated at room temperature for 5min. The diluted DNA and the diluted LIPOFECTAMINE 2000 were mixed together, and the mixture was incubated at room temperature for 30 min. Aspirate the culture medium in the 10cm cell culture dish, add the complex to the cell culture dish, shake the culture plate, and...
Embodiment 2
[0055] 1. Preparation and purification of lentiviral particles
[0056] Take out a certain number of 10cm cell culture dishes, add 4mL of 0.01% poly-lysine to each plate, invert left and right to make the liquid spread on the bottom of the plate, incubate at room temperature for 10min to remove poly-lysine, add 13mL of DMEM to each dish Culture medium (containing 10% FBS) and a certain number of 293t cells were cultured in a cell incubator (37°C, 5% CO 2 ). For each plate of cells, dilute 24 μg DNA with 1.5 mL of serum-free Opti-MEM (pLVTHM:pCMV-dr8.91:pMD2.G plasmid mass ratio is 2:1.5:0.75) and 51 μL with 1.5 mL of serum-free Opti-MEM Transfection reagent Lipofectamine2000, incubated at room temperature for 5min. The diluted DNA and the diluted LIPOFECTAMINE 2000 were mixed together, and the mixture was incubated at room temperature for 30 min. Aspirate the culture medium in the 10cm cell culture dish, add the complex to the cell culture dish, shake the culture plate, and...
Embodiment 3
[0066] 1. Preparation and purification of lentiviral particles
[0067] The virus plasmid pLVTHM was transformed, and the galactosidase gene (LacZ) was inserted, and the successfully transformed plasmid was named pLVTHM-LacZ, and the endotoxin-free large-scale extraction was carried out and reserved for future use. Take out a certain number of 10cm cell culture dishes, add 4mL of 0.01% poly-lysine to each plate, invert left and right to make the liquid spread on the bottom of the plate, incubate for 10min at room temperature to remove poly-lysine, add 13mL of DMEM to each dish Culture medium (containing 10% FBS) and a certain number of 293t cells were cultured in a cell incubator (37°C, 5% CO 2 ). For each plate of cells, dilute 24 μg DNA in 1.5 mL of serum-free Opti-MEM (pLVTHM-LacZ:pCMV-dr8.91:pMD2.G plasmid mass ratio is 2:1.5:0.75), use 1.5 mL of serum-free Opti-MEM Dilute 51 μL of transfection reagent Lipofectamine 2000 and incubate at room temperature for 5 min. The d...
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