G418: Difference between revisions
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==Experimental== | ==Experimental== | ||
* '''stock solution''': 200mg/mL (1000X), stored at -20 C | |||
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. | |||
''neo/kan'' gene is an aminoglycoside 3'-phosphotransferase, which inactivates G418, [[neomycin]] and [[kanamycin]] by phosphorylation. | |||
==Links== | ==Links== | ||
* FAQ on G418 [http://www.g418.com/faq.htm] | * FAQ on G418 [http://www.g418.com/faq.htm] | ||
* list of cell culture material by company [http://www.krackeler.com/products/fid/2910] | |||
Revision as of 06:13, 4 August 2006
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.
