Simple and sensitive procedure for screening yeast mutants that lyse at nonpermissive temperatures.
After mutagenesis, surviving yeast cells are grown on plates at 25 C and later exposed to 37 C. The plates are then overlaid with a soft agar containing p-nitrophenylphosphate at pH 9.7. Lysed cells liberate alkaline phosphatase which gives rise to a yellow color on and around colonies.
D000362:Agar; D000469:Alkaline Phosphatase / Q000378:metabolism; D005020:Ethyl Methanesulfonate; D006863:Hydrogen-Ion Concentration; D009153:Mutagens; D009154:Mutation; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000254:growth & development* / Q000302:isolation & purification; D013696:Temperature
D009153:Mutagens; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D000362:Agar; D005020:Ethyl Methanesulfonate; D000469:Alkaline Phosphatase