What are the recommended concentrations of antibiotics to use for selection in prokaryotes and eukaryotes?
For best results, optimal concentrations for selection should be determined empirically in each unique experiment through dose response curves. However, to get a general idea of concentrations that have worked for individual cell types, please click on the following url: http://www.lifetechnologies.com/us/en/home/life-science/cell-culture/transfection/selection.html or type in Selection Antibiotics into our main search on www.lifetechnologies.com.
Answer Id: 3701
Can Neomycin be used in mammalian selection? Can Neomycin be used instead of Kanamycin in bacterial selection?
No, Neomycin is toxic to mammalian cells. It also causes irreversible damage to kidneys and other organs. Geneticin® (aka G418 Sulfate) is a less toxic and very effective alternative for selection in mammalian cells. Neomycin can be used in bacterial selection, but Kanamycin is the preferred drug to use because of Neomycin's toxicity.
Answer Id: 3949
Can the Neomycin resistance gene found in your mammalian expression vectors also be used for kanamycin selection in E. coli?
No, these vectors will not express kanamycin resistance in E. coli because the Neomycin resistance gene does not have a prokaryotic promoter. You cannot select transformants on kanamycin. This has been verified by streaking a glycerol stock of pcDNA™3.1 vector onto 3 plates: LB, LB + Amp, and LB + Kan. After incubation overnight at 37 degrees C, the cultures were observed to exhibit growth on LB and LB + Amp but no growth on LB + Kan.
Answer Id: 3474