The outcome of this protocol is to produce a solution of G418 and hepes in the concentration of 500 mg of active G418 to 1 ml of hepes.
1. Find the active percentage of G418 as labeled on the vial by the producer (example 760 mg active per g or 76 percent active)
2. Mass the G418 in the vial and calculate the amount active G418 present (example mass of 1.178 mg with 76% active is 1.178*.76 = .89528 g active)
3. Determine the amount of hepes to be used by, dividing the amount of active G418 (in mg) by 500. This is the volume in ml of hepes that needs to be used to generate the desired concentration (example 895.28 mg G418 / 500 = 1.79 ml of hepes)
4. Prepare the solution in a 15 ml tube, filter and aliquot into microtubes in portions of no less the 500 microliters.
5. Store at –20 degree C
Thaw and add G418 at the appropriate concentration. The drug looses activity after one week, make new media with fresh drug as needed.
Vials of G418 can be added to media to the following form to produce the given concentrations for finding the killing curve
for 50 mls of media
200 ug/ml need 20 ul
400 ug/ml need 40 ul
500 ug/ml need 50 ul
600 ug/ml need 60 ul
800 ug/ml need 80 ul
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