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COLLECTION of HUMAN and ANIMAL CELL LINES DSMZ  Form

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[Music] [Applause] after the kill curve and transfection steps we can continue the selection procedure in two different ways depending on the purpose of the developed cell line we can either expand the 24 well to a flask freeze down and start experiments on this pool of resistant cells or we can select further and attempt to isolate a single resistance cell close by limited dilution for selection of a single resistant clone we will need cells detached ing to standard cell culture procedures 10 percent conditioned medium which contains fresh medium with 10 percent old medium to reduce cell stress the selection antibiotic 96-well plates 50 milliliter tubes a pipe header pipettes a multi-channel tips a gutter a laminar flow a microscope and an incubator first we will add 25 milliliters of 10% conditioned selection medium with the concentration of selection antibiotic as determined in the kill curve step 2 3 50 milliliter tubes add the transfected cells to each of the 50 milliliter tubes a

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