Making a long term stock of bacteria - Revision history

Christopher C Vanlang: /* Method */

Christopher C Vanlang — Thu, 28 Jun 2012 08:38:33 GMT

Method

←Older revision		Revision as of 08:38, 28 June 2012
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	==Method==		==Method==
-	*Add 1 ml of 40% glycerol in H<sub>2</sub>O to a cryogenic vial.	+	#Pick a single colony of the clone off a plate and grow an overnight in the appropriate selectable liquid medium (3-5ml).
-	*Add 1 ml sample from the culture of bacteria to be stored.	+	#Add 0.5 ml of 40% glycerol in H<sub>2</sub>O to a cryogenic vial.
-	*Gently vortex the cryogenic vial to ensure the culture and glycerol is well-mixed.	+	#Add 0,5 ml sample from the culture of bacteria to be stored.
-	**Alternatively, pipet to mix.	+	#Gently vortex the cryogenic vial to ensure the culture and glycerol is well-mixed.
-	*Use a tough spot to put the name of the strain or some useful identifier on the top of the vial.	+	#*Alternatively, pipet to mix.
-	*On the side of the vial list all relevant information - part, vector, strain, date, researcher, etc.	+	#On the side of the vial list all relevant information - part, vector, strain, date, researcher, etc.
-	*Store in ~~a freezer box~~ in a -80C freezer. ~~Remember~~ to record ~~where~~ the ~~vial is stored for fast retrieval later~~.	+	#Freeze glycerol stock in liquid nitrogen and store in a -80C freezer.
		+	#*This will also be a good time to record the strain information and record the location.

	==Notes==		==Notes==

Christopher C Vanlang: /* Notes */

Christopher C Vanlang — Thu, 28 Jun 2012 08:34:40 GMT

Notes

←Older revision		Revision as of 08:34, 28 June 2012
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	==Notes==		==Notes==
	*While it is possible to make a long term stock from cells in stationary phase, ideally your culture should be in logarithmic growth phase.		*While it is possible to make a long term stock from cells in stationary phase, ideally your culture should be in logarithmic growth phase.
-		+	*Certain antibiotics in the medium should be removed first as they are supposedly toxic over time, ex)Tetracycline. To do this, spin the culture down and resuspend in same volume of straight LB medium.

	==Specific Protocols==		==Specific Protocols==

Christopher C Vanlang at 08:34, 28 June 2012

Christopher C Vanlang — Thu, 28 Jun 2012 08:34:11 GMT

←Older revision		Revision as of 08:34, 28 June 2012
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	Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria. Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol.		Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria. Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol.

-	==~~Lab~~ Protocols==	+	==Materials==
		+	*40% glycerol solution
		+	* Day/overnight culture
		+	*Cryogenic vials/1.5mL microfuge tube
		+
		+	==Method==
		+	*Add 1 ml of 40% glycerol in H<sub>2</sub>O to a cryogenic vial.
		+	*Add 1 ml sample from the culture of bacteria to be stored.
		+	*Gently vortex the cryogenic vial to ensure the culture and glycerol is well-mixed.
		+	**Alternatively, pipet to mix.
		+	*Use a tough spot to put the name of the strain or some useful identifier on the top of the vial.
		+	*On the side of the vial list all relevant information - part, vector, strain, date, researcher, etc.
		+	*Store in a freezer box in a -80C freezer. Remember to record where the vial is stored for fast retrieval later.
		+
		+	==Notes==
		+	*While it is possible to make a long term stock from cells in stationary phase, ideally your culture should be in logarithmic growth phase.
		+
		+
		+	==Specific Protocols==
	*[[Endy:Making a long term stock of bacteria]]		*[[Endy:Making a long term stock of bacteria]]
	*[[Endy:Retrieving a Registry glycerol]]		*[[Endy:Retrieving a Registry glycerol]]
-		+	*[[-80_Glycerol_Stocks]]
		+	*[[Alm:Glycerol_stocks]]
		+	*[[McClean:E._coli_Glycerol_Stocks]]
		+	*[[Cfrench:glycerol]]
		+	*[[Smolke:Protocols/Freezer_stocks]]
	==Notes==		==Notes==

Torsten Waldminghaus at 17:04, 23 February 2009

Torsten Waldminghaus — Mon, 23 Feb 2009 17:04:52 GMT

←Older revision		Revision as of 17:04, 23 February 2009
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		+	{{back to protocols}}
	==Introduction==		==Introduction==
	Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria. Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol.		Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria. Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol.

Reshma P. Shetty at 21:21, 8 May 2007

Reshma P. Shetty — Tue, 08 May 2007 21:21:59 GMT

←Older revision		Revision as of 21:21, 8 May 2007
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	==Notes==		==Notes==

-	[[Category:Protocol]] [[Category:Escherichia coli]]	+	[[Category:Protocol]]
		+	[[Category:In vivo]]
		+	[[Category:Escherichia coli]]

Reshma P. Shetty at 19:30, 27 April 2007

Reshma P. Shetty — Fri, 27 Apr 2007 19:30:44 GMT

←Older revision		Revision as of 19:30, 27 April 2007
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	==Lab Protocols==		==Lab Protocols==
	*[[Endy:Making a long term stock of bacteria]]		*[[Endy:Making a long term stock of bacteria]]
-	*[[Endy:Retrieving a Registry ~~glycerol\|Retrieving a registry~~ glycerol]]	+	*[[Endy:Retrieving a Registry glycerol]]

	==Notes==		==Notes==
		+
		+	[[Category:Protocol]] [[Category:Escherichia coli]]

Josh K. Michener: /* Lab Protocols */

Josh K. Michener — Wed, 25 Jan 2006 20:23:48 GMT

Lab Protocols

←Older revision		Revision as of 20:23, 25 January 2006
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	==Lab Protocols==		==Lab Protocols==
-	[[Endy:Making a long term stock of bacteria]]	+	*[[Endy:Making a long term stock of bacteria]]
		+	*[[Endy:Retrieving a Registry glycerol\|Retrieving a registry glycerol]]

	==Notes==		==Notes==

Barry Canton at 15:52, 20 January 2006

Barry Canton — Fri, 20 Jan 2006 15:52:43 GMT

New page

==Introduction==
Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria. Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol.

==Lab Protocols==
[[Endy:Making a long term stock of bacteria]]

==Notes==