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

Karmella Haynes: /* 02/15/13 */

2013-02-19T01:41:17Z

02/15/13

←Older revision		Revision as of 01:41, 19 February 2013
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	# 6-well plate: 2 mL of 1:25 dln., ~80,000 cells/ well		# 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		# 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 ~~(Note~~: ~~was~~ careful to gently mix stock tube by inverting after every column was filled to ensure that cells stayed suspended in the stock tube)	+	# 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.

Karmella Haynes: /* 02/15/13 */

2013-02-19T01:40:07Z

02/15/13

←Older revision		Revision as of 01:40, 19 February 2013
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	**(1) 1:25 dilution of harvested cells (1mL cells + 24 mL medium) = 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 previous tube (1mL 1:25 cells + 24 mL medium) = 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 harvested cells --> 1:100 dilution of these	+	**(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)

		+
		+	Plates:
	# 6-well plate: 2 mL of 1:25 dln., ~80,000 cells/ well		# 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		# 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	+	# 96-well plate: 200 uL of 1:40,000 dln., ~5 cells/ well (Note: was careful to gently mix stock tube by inverting after every column was filled to ensure that cells stayed suspended in the stock tube)

Karmella Haynes: /* 02/15/13 */

2013-02-19T01:34:11Z

02/15/13

←Older revision		Revision as of 01:34, 19 February 2013
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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 ~~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

Karmella Haynes: /* Entry title */

2013-02-19T01:30:31Z

Entry title

←Older revision		Revision as of 01:30, 19 February 2013
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	\| colspan="2"\|		\| colspan="2"\|
	<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
-	==~~Entry title~~==	+	==02/15/13==
-	* ~~Insert content here...~~	+	* HEK293-luc 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
		+	* Retry 6-well plates and also try 96-well plate
		+	* Made three new plates of HEK293-luc #4 cells
		+	* Started with ~1x10<sup>6</sup> harvested cells (confluent T25 flask)
		+
		+	* Made the following dilutions:
		+	**(1) 1:25 dilution of harvested cells = 40,000 cells/mL 10% FBS DMEM
		+	**(2) 1:25 dilution of tube 1 = 1:625 dilution = 1,600 cells/mL 10% FBS DMEM
		+	**(3) 1:100 dilution of 1x10<sup>6 cells/mL
		+
		+	# 6-well plate: 2 mL , 80,000 cells/ well
		+	# 6-well plate: 2 mL 10% FBS DMEM, 3200 cells/ well
		+

Karmella Haynes: Autocreate 2013/02/15 Entry for User:David_Dreher/Notebook/Chromatin_controlled_cell_pattern

2013-02-19T01:19:46Z

Autocreate 2013/02/15 Entry for User:David_Dreher/Notebook/Chromatin_controlled_cell_pattern

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