View source for Yu:Lysis

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|+'''Quick Yeast Lysis Procedure'''
|width="600px" valign="top"|1. Grow yeast cells to late-log phase
|width="200px" valign="top"|Grow yeast
|-
|2. Pellet 2mL cell culture in fast-prep tube. Remove excess supernatant.
|2mL yeast culture pelleted in fastprep tube
|-
|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.
|valign="top"|Prep completed lysis buffer per tube:
|-
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{| border="0" cellpadding="5" style="width:50%"
|+ '''Completed <span id="buffer">lysis buffer</span>'''
|align="left"|[[Yu:PBS#PBSMT|PBSMT]]
|-
|5μL 200mM [[PMSF]] per 1mL PBSMT 
|-
|2.6μL .5M Benzamide per 1mL PBSMT
|-
|1μL [[Yu:PLAAC|PLAAC]] per 1mL PBSMT
|}
|-
|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
|-
|5. Add approximately 100μL acid-washed glass fastprep beads to each tube.
|100μL beads per tube
|-
|6. Fastprep at 6.5 m/s for 30 seconds
|Fastprep at 6.5 for 30s
|-
|7. Add 500μL completed lysis buffer to each fastprep tube
|500μL comp. lysis buffer per tube
|-
|8. Spin at top speed for 10 minutes at 4°C
|Spin top speed 10 minutes at 4°C
|-
|9. Pipette off supernatant, which contains lysates. Store on ice or in -20°C freezer.
|Remove and save supernatant
|}