G418
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Experimental
- stock solution: 200mg/mL (1000X), stored at -20 C
- working concentration: working concentration depends on organism and purpose of antibiotic application; stable at RT
- bacteria selection: 8-16 µg/ml
- plant cell maintenance: 10 µg/ml
- plant cell selection: 25-50 µg/ml
- mammalian cell maintenance: 200 µg/ml
- mammalian cell selection: 400 µg/ml (range: 300-1000 µg/ml)
- yeast: after transformation incubate culture at 30°C for 2-3 hours and then plate on YPD containing 200 mg/L (=µg/ml) G418.
Description
- Gentamycin analog, used to select for the KanMX marker in yeast.
- mode of action: irreversibly binds to 80S ribosomal subunit, disrupting proofreading.
- mode of resistance: neo/kan gene is an aminoglycoside 3'-phosphotransferase, which inactivates G418, neomycin and kanamycin by phosphorylation.
