Antibiotic kill curve is a dose response experiment in which mammalian cells are subjected to increasing amounts of selection antibiotic to determine the minimum concentration of an antibiotic that can kill all the cells in a specific period of time. This is a crucial step before using a selection antibiotic to kill untransfected cells and generate stable cell lines. Since each mammalian cell line differs in antibiotic sensitivity, it is always recommended to perform separate kill curve experiments for different cell lines and antibiotics.
The table below indicates the concentration ranges for commonly used selection antibiotics that may be adopted in dose response experiments.
The dose response may vary with the cell line being tested. The concentration ranges are only for guidance; a kill curve must be performed with every selection antibiotic when used for the first time or used on a new cell line.
The following protocol outlines the steps to generate kill curve for your cells.
What you need:
Note: Antibiotics are most effective when the cells are in active division; an optimum cell density is therefore recommended for most accurate dose response.
Figure 1.Schematic guide – selection antibiotic concentration.
Figure 2.Mock Antibiotic Kill Curve
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