Haynes Lab:Notebook/Short Projects/2014/09/10: Difference between revisions

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'''Cell culture'''
'''Cell culture'''
* Set up 2x 6-well plates for Gal4-VP64 transfections
* Set up 2x 6-well plates for Gal4-VP64 transfections
** 6 wells dox- (3 will be transfected, 3 not)
** 6 wells dox- (3 will be transfected, 3 not) --> 2.5E/well
** 6 wells dox+ (3 will be transfected, 3 not)
** 6 wells dox+ (3 will be transfected, 3 not) --> 2.5E/well
# Follow steps 1-9 of [http://openwetware.org/wiki/Haynes:SplittingCells Passaging Adherent Cells] protocol.
<br>Note: cells are ~75% confluent, so at this point you will have 10 mL of cells (~7.5E5/ mL).
#
 


* Also, passage cells 1:5 in T-75 flasks
* Also, passage cells 1:5 in T-75 flasks

Revision as of 09:02, 10 September 2014

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09/10/14 - Jonah Thomas

  • Cell culture - Gal4-EED/luc for luciferase activation experiment
  • Passage cells 1:5 (T-75 flasks)


Cell culture

  • Set up 2x 6-well plates for Gal4-VP64 transfections
    • 6 wells dox- (3 will be transfected, 3 not) --> 2.5E/well
    • 6 wells dox+ (3 will be transfected, 3 not) --> 2.5E/well
  1. Follow steps 1-9 of Passaging Adherent Cells protocol.


Note: cells are ~75% confluent, so at this point you will have 10 mL of cells (~7.5E5/ mL).


  • Also, passage cells 1:5 in T-75 flasks
  • Make fresh growth medium: DMEM, 10% FBS, 1% pen-strep



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