User:David Dreher/Notebook/Chromatin controlled cell pattern/2013/02/15

(Difference between revisions)

Revision as of 21:34, 18 February 2013

Project name

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
Retry 6-well plates and also try 96-well plate
Made three new plates of HEK293-luc #4 cells
Started with ~1x10⁶ harvested cells (confluent T25 flask)

Made the following dilutions:
- (1) 1:25 dilution of harvested cells (1mL cells + 24 mL medium) = 40,000 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 harvested cells --> 1:100 dilution of these

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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
@@ Line 19: / Line 19: @@
 * 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 1x10<sup>6 cells/mL
+**(3) 1:100 dilution of harvested cells --> 1:100 dilution of these
-# 6-well plate: 2 mL , 80,000 cells/ well
+# 6-well plate: 2 mL of 1:25 dln., ~80,000 cells/ well
-# 6-well plate: 2 mL 10% FBS DMEM, 3200 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