G418: Difference between revisions

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Used to select for the KanMX marker in yeast.
[[Aminoglycoside]] antibiotic. G418/G-418 (generic name) is the same molecule as Geneticin® (registered trademark of Invitrogen Corp.).


After transformation incubate culture at 30 C for 2-3 hours and then plate on [[YPD]] containing 200 mg/L G418.
==Experimental==


The stock solution is 200mg/mL (1000X) and is stored at -20 C
* '''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.
 
==Links==
* related antibiotics: [[kanamycin]], [[neomycin]] (all aminoglycosides)
* FAQ on G418 [http://www.g418.com/faq.htm]
* list of cell culture material by company [http://www.krackeler.com/products/fid/2910]
 
[[Category:Material]] [[Category:Antibiotic]]

Latest revision as of 10:46, 26 July 2007

Aminoglycoside antibiotic. G418/G-418 (generic name) is the same molecule as Geneticin® (registered trademark of Invitrogen Corp.).

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.

Links

  • related antibiotics: kanamycin, neomycin (all aminoglycosides)
  • FAQ on G418 [1]
  • list of cell culture material by company [2]