Quint Lab:yeast plasmid prep

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yeast plasmid prep inoculate 2 ml of Glucose medium and let grow over night @ 30°C spin down 1.5 ml of the over night culture and resuspend in0.5 ml S-buffer incubate 30 min, 37°C add 0.1 ml lysis solution, vortex incubate 30 min, 65°C add 0.166 ml potassium acetate and mix, chill on ice for 10 min spin down for 10 min, pour the SN into a new tube (2 ml!) DNA precipitation: add 0.8 ml EtOH, 10 min on ice, spin down for 10 min, pour off the SN wash DNA with 0.5 ml 70% EtOH, dry pellet dissolve DNA in 0.04 ml water S-buffer: 10 mM K2HPO4; pH 7.2 10 mM EDTA 50 mM 2-Mercaptoethanol 50 µg/ml zymolase lysis solution: 25 mM Tris-HCl; pH 7.5 25 mM EDTA 2.5% SDS