Knight:Centrifuge desalting/Zeba columns - Revision history

Reshma P. Shetty at 22:37, 5 December 2006

2006-12-05T22:37:54Z

←Older revision		Revision as of 22:37, 5 December 2006
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	#Centrifuge at 1500 x ''g'' for 1 minute.		#Centrifuge at 1500 x ''g'' for 1 minute.
	#Discard desalting column.		#Discard desalting column.
		+
		+	[[Category:Protocol]]
		+	[[Category:In vitro]]
		+	[[Category:Protein]]

Reshma P. Shetty at 21:43, 27 November 2006

2006-11-27T21:43:35Z

←Older revision		Revision as of 21:43, 27 November 2006
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-	~~<font color=red>in progress!</font>~~
-
	==Overview==		==Overview==
	This protocol is for doing buffer exchange on small sample volumes (30-130 μL) volumes.		This protocol is for doing buffer exchange on small sample volumes (30-130 μL) volumes.

	==Materials==		==Materials==
-	*Zeba 0.~~5mL desalting spin columns~~ from Pierce	+	*[http://www.piercenet.com/products/browse.cfm?fldID=2845ECFE-7622-4D7B-B114-73C6D523402D Zeba Desalt Spin Columns, 0.5 ml] from Pierce
	*Variable-speed bench-top microcentrifuge		*Variable-speed bench-top microcentrifuge
	*2.0 mL collection tubes		*2.0 mL collection tubes

Reshma P. Shetty at 14:42, 16 October 2006

2006-10-16T14:42:46Z

←Older revision		Revision as of 14:42, 16 October 2006
Line 1:		Line 1:
		+	<font color=red>in progress!</font>
		+
	==Overview==		==Overview==
	This protocol is for doing buffer exchange on small sample volumes (30-130 μL) volumes.		This protocol is for doing buffer exchange on small sample volumes (30-130 μL) volumes.

Reshma P. Shetty at 14:39, 16 October 2006

2006-10-16T14:39:13Z

New page

==Overview==
This protocol is for doing buffer exchange on small sample volumes (30-130 μL) volumes.

==Materials==
*Zeba 0.5mL desalting spin columns from Pierce
*Variable-speed bench-top microcentrifuge
*2.0 mL collection tubes
*[[Knight:Protein DNA binding|Buffer]] for exchange

==Procedure==
#Remove column's bottom closure.
#Loosen cap.
#Place column in a 2.0 mL microcentrifuge collection tube.
#Centrifuge at 1500 x ''g'' for 1 minute.
#*Removes storage solution.
#Discard storage solution.
#Mark orientation of column in centrifuge so that the column is placed in the same orientation each time.
#Add 300 μL [[Knight:Protein DNA binding|Buffer]].
#Centrifuge at 1500 x ''g'' for 1 minute.
#Discard buffer.
#Add 300 μL [[Knight:Protein DNA binding|Buffer]].
#Centrifuge at 1500 x ''g'' for 1 minute.
#Discard buffer.
#Add 300 μL [[Knight:Protein DNA binding|Buffer]].
#Centrifuge at 1500 x ''g'' for 1 minute.
#Discard buffer.
#Add 300 μL [[Knight:Protein DNA binding|Buffer]].
#Centrifuge at 1500 x ''g'' for 1 minute.
#Discard buffer.
#Move column to new 2.0 mL collection tube.
#Apply 30-130 μL of sample to top of compact resin bed.
#*''For volumes less than 70 μL, apply 15 μL stacker of ultrapure water or buffer to top of compact resin bed after sample has absorbed to improve sample recovery.''
#Centrifuge at 1500 x ''g'' for 1 minute.
#Discard desalting column.