Difference between revisions of "Phleomycin"

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Used to select for the ble marker in yeast.
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==Mode of Action==
 
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Binds and cleaves DNA
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==Mechanism of Resistance==
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The product of the ''Sh ble'' gene binds to phleomycin and inhibits its ability to cleave DNA
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==Usage==
 
After transformation incubate culture in YPD at 30 C for 6 hours to overnight, then chill on ice for 1 hour. Plate on YPD (pH 7) containing 10 mg/L phleomycin.
 
After transformation incubate culture in YPD at 30 C for 6 hours to overnight, then chill on ice for 1 hour. Plate on YPD (pH 7) containing 10 mg/L phleomycin.
  
 
Phleomycin is pH sensitive--it loses its effect at low pH. When making phleomycin plates add 2 pellets of NaOH per liter to get the pH above 7.
 
Phleomycin is pH sensitive--it loses its effect at low pH. When making phleomycin plates add 2 pellets of NaOH per liter to get the pH above 7.
  
The stock solution is 10 ug/mL (1000X), is light blue and is stored at -20 C. It should be stable for 1 year.
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The stock solution is 20 ug/mL (2000X), is light blue and is stored at -20 C. It should be stable for 1 year.
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The best price I've found is through Invivogen
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[[Category:Material]] [[Category:Antibiotic]]

Latest revision as of 10:47, 26 July 2007

Mode of Action

Binds and cleaves DNA

Mechanism of Resistance

The product of the Sh ble gene binds to phleomycin and inhibits its ability to cleave DNA

Usage

After transformation incubate culture in YPD at 30 C for 6 hours to overnight, then chill on ice for 1 hour. Plate on YPD (pH 7) containing 10 mg/L phleomycin.

Phleomycin is pH sensitive--it loses its effect at low pH. When making phleomycin plates add 2 pellets of NaOH per liter to get the pH above 7.

The stock solution is 20 ug/mL (2000X), is light blue and is stored at -20 C. It should be stable for 1 year.

The best price I've found is through Invivogen