User:Daniel Goodman/Notebook/Cluzel/2010/03/31: Difference between revisions
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Protocol for Competent Cells | |||
*Grow 250 ml flask, one colony, 50 ml LB at 37 deg for 4 hours (OD must be 0.3-0.4) in shaker (check at 7pm) | |||
*Spin down 20 mls in 2 50 ml epis at 4 degrees in centrifuge | |||
**3200 RCF (4000 RPM) 10 minutes 4 degrees | |||
** pour of supernatant, add 40 mls 4 deg filtered water each (maybe add 5 first, vortex, add 35 more) | |||
*** make sure pellet is dissolved | |||
** repeat above 2x | |||
* resuspend pellets in 10% glycerol (5 mls each) | |||
* spin, remove glycerol supernatant | |||
* resuspend in 0.5 ml 10% glycerol each, combine tubes | |||
* make 20x 50 ul aliquots, put in -80 freezer | |||
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Revision as of 12:10, 31 March 2010
Cluzel Lab Notebook Daniel Goodman |
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Entry title
Protocol for Competent Cells
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