Quint Lab:yeast plasmid prep

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yeast plasmid prep

 * 1) inoculate 2 ml of Glucose medium and let grow over night @ 30°C
 * 2) spin down 1.5 ml of the over night culture and resuspend in0.5 ml S-buffer
 * 3) incubate 30 min, 37°C
 * 4) add 0.1 ml lysis solution, vortex
 * 5) incubate 30 min, 65°C
 * 6) add 0.166 ml potassium acetate and mix, chill on ice for 10 min
 * 7) spin down for 10 min, pour the SN into a new tube (2 ml!)
 * 8) DNA precipitation: add 0.8 ml EtOH, 10 min on ice, spin down for 10 min, pour off the SN
 * 9) wash DNA with 0.5 ml 70% EtOH, dry pellet
 * 10) 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