G418 selection of stable cell

oBWhat · #1 (**permalink**) 07-07-2006, 08:42 PM

How can I conduct a G418 selection to select for stable cell lines? I am trying to integrate a plasmid sequence containing G418 resistance and a promoter driving the expression of a gene X.

What is the protocol / general procedure for this? any tips to do this? thanks

bioinfofreak · #2 (**permalink**) 08-08-2006, 07:56 PM

You need to get your hands on some information first. See what pubmed articles did G418 selection for your cell line of interest. what concentrations did they use? how long they selected?

using these you can figure out a few things... then just make a concentration curve and find your LD50 that kills 50% of your cells... after a certain time you can get selected cells growing. these should have your vector of interest integrated.

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G418 selection of stable cell

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