CATHY'S NO BOIL YEAST EXTRACTS
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CATHY'S NO BOIL YEAST EXTRACTS
1.Cells to mid log/ need ~ 5OD.Spin down and wash w/ 1ml H2O
2.Transfer to 1.5ml round bottom screw cap tube
3.add 200μl SDS sample buffer+ (5% BME,50mM NaF,1mM PMSF)-add to stock just before use; + 200μl glass beads
4.Beat 90sec (setting 9)in cold room and spin down 2',4℃.
Sample buffer: 4% SDS 5ml+
80mM Tris Cl6.8
Final 10% glycerol
10mM EDTA
0.2% BFB