Quick Yeast Lysis Procedure
| 1. Grow yeast cells to late-log phase
| Grow yeast
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| 2. Pellet 2mL cell culture in fast-prep tube. Remove excess supernatant.
| 2mL yeast culture pelleted in fastprep tube
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| 3. Prepare 600μL completed yeast lysis buffer per fastprep tube. Multiply each ingredient by the number of fastprep tubes to be lysed. Keep on ice after prepared and while using.
| Prep completed lysis buffer per tube:
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Completed lysis buffer
| PBSMT
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| 5μL 200mM PMSF per 1mL PBSMT
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| 2.6μL .5M Benzamide per 1mL PBSMT
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| 1μL PLAAC per 1mL PBSMT
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| 4. Add 100μL complete lysis buffer to each fastprep tube. Keep on ice or in cold room for remaining steps.
| 100μL comp. lysis buffer per tube
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| 5. Add approximately 100μL acid-washed glass fastprep beads to each tube.
| 100μL beads per tube
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| 6. Fastprep at 6.5 m/s for 30 seconds
| Fastprep at 6.5 for 30s
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| 7. Add 500μL completed lysis buffer to each fastprep tube
| 500μL comp. lysis buffer per tube
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| 8. Spin at top speed for 10 minutes at 4°C
| Spin top speed 10 minutes at 4°C
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| 9. Pipette off supernatant, which contains lysates. Store on ice or in -20°C freezer.
| Remove and save supernatant
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