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Tuesday, November 18, 2003
Description Yeast Glass Bead Prep Procedure Sample Buffer: 0.06M Tris-HCl, pH 6.8 10% (v/v) glycerol 2% (w/v) SDS 5% (v/v) 2-mercaptoethanol 0.0025% (w/v) bromophenol blue 0.1 M PMSF 0.5 M Benzamidine 1. Grow 25 ml of cells to mid-log. 2. Spin down (2500 rpm for 5 minutes). Wash 1X with water and spin again. 3. Resuspend in 1 ml of water and transfer to 1.5 ml microfuge tube. Spin down for 5 seconds and pour off water. 4. Resuspend in 0.5 ml of ice cold Sample Buffer with freshly added PMSF (0.5 mM) and benzamidine (0.5 mM). 5. Add glass beads (~0.5 ml). 6. Vortex on high 4X for 45 seconds with 30 seconds on ice in between each mixing. 7. Spin for 5 minutes in microfuge at 4 deg C. 8. Transfer supernatant to a new tube and boil for 5 minutes. 9. Load 10-15 l on a protein gel. Recipes Supplies Tips (责任编辑：泉水) |
Yeast Glass Bead Prep
时间:2005-07-18 00:00来源:Scienceboard.net 作者:admin 点击:
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