G418

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(working concentrations by organism)
(resistance info added, links to neo & kan)
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* mammalian cell selection: 400 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.
* yeast: after transformation incubate culture at 30°C for 2-3 hours and then plate on [[YPD]] containing 200 mg/L (=µg/ml) G418.
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==Mode of Action==
==Mode of Action==
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+
 
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.
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 +
==Mode of resistance==
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 +
''neo/kan'' gene is an aminoglycoside 3'-phosphotransferase, which inactivates G418, [[neomycin]] and [[kanamycin]] by phosphorylation.
==Safety==
==Safety==

Revision as of 08:51, 4 August 2006

Contents

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]
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