Use of Petri Dish in the Screening and Cloning of Transfected Cells

Use of Petri Dish in the Screening and Cloning of Transfected Cells

1. 48 hours after transfection, the cells were divided into a culture medium containing 125 μmol/L L-histidine and 0.5 μg/ml tetracycline-HCl using several dilutions. The cell density is approximately 1 × 106 to 3 × 104 per 10 cm culture dish. A petri dish containing approximately 1 × 105 cells needs to be cultured in several.medical petri dish,round petri dish,Wholesale Petri Dish,petri dish disposable
2. After the cells are cultured in the selection medium for 4 days, they are then cultured with 3 to 4 ml of the selection medium containing 125 μmol/L L-histidine and 0.5 μg/ml tetracycline-HCl.
3. After the colony is formed (usually after about 10-12 days of selective culture), replace it with a selective culture medium containing 250 μmol/L L-histidine and 0.5 μg/ml tetracycline-HCl.
4. After the colonies have grown (choose to cultivate for 12 to 15 days), draw the boundaries of each colony with a circle on the bottom of the petri dish. Aspirate the culture medium and place a sterile plastic cloning ring on the individual clones in the petri dish. Repeat this step for each selected colony. To select cells from a culture dish, individual colonies are required to be relatively separated and easy to distinguish.medical petri dish,round petri dish,Wholesale Petri Dish,petri dish disposable
5. Wash the clone quickly with about 100 μl phosphate buffer. In order to make the cells fall off, add 2 drops of 1 × pancreatin-EDTA (100 μl) and leave it for 30-60s. Pipette up and down with a Pasteur pipette to loosen the cells. Each colony was transferred to a well of a 24-well tissue culture medium plate, and each well contained 1 ml of selective medium containing 250 μmol/L L-histidine and tetracycline.
6. When the cells in the wells reach 80% confluence, transfer them to a 6 cm tissue culture dish, and use a selective culture medium containing 500 μmol/L L-histidine and tetracycline.
7. Cells are expanded and cultured in a selective culture medium containing 500 μmol/L L-histidine and tetracycline (usually the cells are diluted 1: 5 to 1: 10).medical petri dish,round petri dish,Wholesale Petri Dish,petri dish disposable