G418: Difference between revisions
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(working concentrations by organism) |
(resistance info added, links to neo & kan) |
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* yeast: after transformation incubate culture at 30°C for 2-3 hours and then plate on [[YPD]] containing 200 mg/L (=µg/ml) G418. | * yeast: after transformation incubate culture at 30°C for 2-3 hours and then plate on [[YPD]] containing 200 mg/L (=µg/ml) G418. | ||
==Mode of Action== | |||
Irreversibly binds to 80S ribosomal subunit, disrupting proofreading. Can be blocked by aminoglycoside-3'-phosphotransferase. | Irreversibly binds to 80S ribosomal subunit, disrupting proofreading. Can be blocked by aminoglycoside-3'-phosphotransferase. | ||
==Mode of resistance== | |||
''neo/kan'' gene is an aminoglycoside 3'-phosphotransferase, which inactivates G418, [[neomycin]] and [[kanamycin]] by phosphorylation. | |||
==Safety== | ==Safety== |
Revision as of 05:51, 4 August 2006
Experimental
Gentamycin analog, used to select for the KanMX marker in yeast.
Stock Solution
200mg/mL (1000X), stored at -20 C
Working Concentration
Working concentration depends on organism and purpose of antibiotic application.
- 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
- yeast: after transformation incubate culture at 30°C for 2-3 hours and then plate on YPD containing 200 mg/L (=µg/ml) G418.
Mode of Action
Irreversibly binds to 80S ribosomal subunit, disrupting proofreading. Can be blocked by aminoglycoside-3'-phosphotransferase.
Mode of resistance
neo/kan gene is an aminoglycoside 3'-phosphotransferase, which inactivates G418, neomycin and kanamycin by phosphorylation.
Safety
Links
- FAQ on G418 [1]