User:David Dreher/Notebook/Chromatin controlled cell pattern/2013/02/15: Difference between revisions
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''HEK293-luc #4 cell culturing, trial 2'' | '''HEK293-luc #4 cell culturing, trial 2''' | ||
* Tried plating at the optimal density (1600 cells/well) in a 6-well plate; cells did not grow very well | * Tried plating at the optimal density (1600 cells/well) in a 6-well plate; cells did not grow very well | ||
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* Made the following dilutions: | * Made the following dilutions: | ||
**(1) 1:25 dilution of harvested cells = 40,000 cells/mL 10% FBS DMEM | **(1) 1:25 dilution of harvested cells (1mL cells + 24 mL medium) = 40,000 cells/mL 10% FBS DMEM | ||
**(2) 1:25 dilution of tube 1 = 1:625 dilution = 1,600 cells/mL 10% FBS DMEM | **(2) 1:25 dilution of previous tube (1mL 1:25 cells + 24 mL medium) = 1:625 dilution = 1,600 cells/mL 10% FBS DMEM | ||
**(3) 1:100 dilution of | **(3) 1:100 dilution of harvested cells (100 μL cells + 9.9 mL medium) <br>--> 1:100 dilution of these (100 μL cells + 9.9 mL '''20% FBS''' DMEM) <br>--> 1:4 dilution of these (5 mL cells + 15 mL '''20% FBS''' DMEM) <br>= 25 cells/mL = 5 cells/200 μL (20 mL total) | ||
# 6-well plate: 2 mL , 80,000 cells/ well | |||
# 6-well plate: 2 mL | Plates: | ||
# 6-well plate: 2 mL of 1:25 dln., ~80,000 cells/ well | |||
# 6-well plate: 2 mL of 1:625 dln., ~3,200 cells/ well | |||
# 96-well plate: 200 uL of 1:40,000 dln., ~5 cells/ well | |||
Notes: | |||
* Was careful to gently mix stock tube by inverting after every column was filled to ensure that cells stayed suspended in the stock tube. | |||
* Used 20% FBS for 96-well plate to encourage growth of isolated/ low-concentration cells. | |||
Revision as of 18:41, 18 February 2013
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02/15/13
HEK293-luc #4 cell culturing, trial 2
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