Screening method for hybrid tumor cell monoclonal preparation
A technology for hybridoma cells and screening methods, applied in biochemical equipment and methods, microbial determination/inspection, fermentation, etc., can solve the source and quality limit of fluorescent latex microbeads, uneven cell distribution, and difficult cell mixing, etc. It can reduce the risk of pollution, increase the number of clones, and save the tedious operation.
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Embodiment 1
[0021] (1) Preparation of 2 times DMEM basal culture solution: Take 13.4 grams of DMEM, add 500 milliliters of pure water, add 3.8 grams of sodium bicarbonate and fully stir to dissolve, adjust the pH to 7.2. Sterilize by filtration to prepare 2 times DMEM basal culture solution.
[0022] (2) Preparation of L-glutamine mother liquor: Weigh 5.8 grams of L-glutamine (L-Glutamine, Invitrogen), add 200 milliliters of pure water, heat to 50 ° C to completely dissolve, sterilize and filter to obtain 200 mmol For every liter of L-glutamine mother solution, divide into 4ml per tube and store at -20°C.
[0023] (3) Preparation of β-mercaptoethanol mother liquor: Take 70 microliters of β-mercaptoethanol, add it to 100 milliliters of pure water, filter and sterilize to obtain 10 millimoles per liter of β-mercaptoethanol mother liquor, and store at -20°C.
[0024] (4) Preparation of methyl cellulose aqueous solution: take 0.6 g of 4000 cp methyl cellulose and 25 ml of ultrapure water and...
Embodiment 2
[0029] (1) Preparation of 2 times DMEM basal culture solution: Take 13.4 grams of DMEM, add 500 milliliters of pure water, add 3.8 grams of sodium bicarbonate and fully stir to dissolve, adjust the pH to 7.4. Sterilize by filtration to prepare 2 times DMEM basal culture solution.
[0030] (2) Preparation of L-glutamine mother liquor: Weigh 2.9 grams of L-glutamine (L-Glutamine, Invitrogen), add 100 milliliters of pure water, heat to 52 ° C to completely dissolve, sterilize and filter to obtain 200 mmol For every liter of L-glutamine mother solution, divide into 4ml per tube and store at -20°C.
[0031] (3) Preparation of β-mercaptoethanol mother solution: Take 63 microliters of β-mercaptoethanol, add it to 90 ml of aqueous solution, filter and sterilize to obtain 10 mmol per liter of β-mercaptoethanol mother solution, and store at -20°C.
[0032] (4) Preparation of methyl cellulose aqueous solution: take 0.6 g of 4000 cp methyl cellulose and 25 ml of pure water and sterilize at...
Embodiment 3
[0037] (1) Preparation of 2 times DMEM basal culture solution: Take 13.4 grams of DMEM, add 500 milliliters of pure water, add 3.8 grams of sodium bicarbonate and fully stir to dissolve, adjust the pH to 7.2. Sterilize by filtration to prepare 2 times DMEM basal culture solution.
[0038] (2) Preparation of L-glutamine mother liquor: Weigh 8.7 grams of L-glutamine (L-Glutamine, Invitrogen), add 150 ml of pure water, heat to 50°C to completely dissolve, sterilize and filter to obtain 200 mmol For every liter of L-glutamine mother solution, divide into 4ml per tube and store at -20°C.
[0039] (3) Preparation of β-mercaptoethanol mother solution: Take 77 microliters of β-mercaptoethanol, add it to 110 ml of aqueous solution, filter and sterilize to obtain 10 mmol per liter of β-mercaptoethanol mother solution, and store at -20°C.
[0040] (4) Preparation of methyl cellulose aqueous solution: Take 0.9 g of 4000 cp methyl cellulose and 25 ml of pure water and sterilize at 121-126...
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