Team:British Columbia/Notebook DspB

From 2010.igem.org

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Line 13: Line 13:
<li>Add 850uL of TE or EB buffer (fill up to 1mL)</li>
<li>Add 850uL of TE or EB buffer (fill up to 1mL)</li>
<li>Sonicate (ear protectors on!)</li>
<li>Sonicate (ear protectors on!)</li>
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<ol>
+
<ol type="i">
<li>Turn sonicator on</li>
<li>Turn sonicator on</li>
<li>Wipe down probe with kimwipe wetted with ethanol</li>
<li>Wipe down probe with kimwipe wetted with ethanol</li>

Revision as of 04:41, 27 October 2010


Sonication Protocol to Lyse Cells

  1. Add 100uL of sdH2O to 5mg of lysozyme
  2. Pipet this into the cell pellet and resuspend
  3. Add 50uL of 0.5M EDTA pH 8.0 to a final concentration of 25uM
  4. Add 850uL of TE or EB buffer (fill up to 1mL)
  5. Sonicate (ear protectors on!)
    1. Turn sonicator on
    2. Wipe down probe with kimwipe wetted with ethanol
    3. Press start
    4. Turn knob up to desired level (in this case, 5)
    5. Place mirocentrifuge tube under the probe and move it up and down for 15 seconds
    6. Take the tube out and place on ice
    7. Repeat 5 more times
    8. Turn off sonicator




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