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
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 10 ug/mL (1000X), is light blue and is stored at -20 C. It should be stable for 1 year.