Endy:STN Chemostat Protocol - Revision history

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

2007-04-27T19:27:30Z

←Older revision		Revision as of 19:27, 27 April 2007
Line 22:		Line 22:
	*Mark 18 mL, 20 mL, and 22 mL on chemostat chambers to assist in monitoring volume.		*Mark 18 mL, 20 mL, and 22 mL on chemostat chambers to assist in monitoring volume.

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

Reshma P. Shetty at 19:26, 27 April 2007

2007-04-27T19:26:55Z

←Older revision		Revision as of 19:26, 27 April 2007
Line 21:		Line 21:
	*Measure the flow rate by the effluent. 10 rpm gives about 0.25 mL/min, which is about right, but flow rate should be checked during each run.		*Measure the flow rate by the effluent. 10 rpm gives about 0.25 mL/min, which is about right, but flow rate should be checked during each run.
	*Mark 18 mL, 20 mL, and 22 mL on chemostat chambers to assist in monitoring volume.		*Mark 18 mL, 20 mL, and 22 mL on chemostat chambers to assist in monitoring volume.
		+
		+	[[Category:Escherichia coli]]

Cconboy at 17:29, 20 June 2006

2006-06-20T17:29:21Z

←Older revision		Revision as of 17:29, 20 June 2006
Line 1:		Line 1:
	contact: [[Caitlin Conboy]], [[Jennifer Braff]]		contact: [[Caitlin Conboy]], [[Jennifer Braff]]
		+	===Chemostat Design===
		+
		+	See [[Endy:STN Chemostat Design\|Chemostat Design]].
		+
		+	===Chemostat Protocol===
	==Day 0==		==Day 0==
	Set up o/n culture in supM9 media in a small flask at 37 C.		Set up o/n culture in supM9 media in a small flask at 37 C.
-

	==Day 1==		==Day 1==
Line 10:		Line 14:
	*Add the media input (teal, 23-G1 needle) and turn on pump (currently 10 rpm for 20 mL/chamber). After about 1 hour, check that the culture volume is 20 mL and adjust effluent as necessary.		*Add the media input (teal, 23-G1 needle) and turn on pump (currently 10 rpm for 20 mL/chamber). After about 1 hour, check that the culture volume is 20 mL and adjust effluent as necessary.
	*The cultures should wash out to a stable OD600 by the morning (day 2). OD600 should be monitored throughout the day (~every 3 hrs) and samples drawn for steady-state measurement either in the latter part of day 2 or on day 3.		*The cultures should wash out to a stable OD600 by the morning (day 2). OD600 should be monitored throughout the day (~every 3 hrs) and samples drawn for steady-state measurement either in the latter part of day 2 or on day 3.
-

	==Notes==		==Notes==

Cconboy: Endy: STN Chemostat Protocol moved to Endy:STN Chemostat Protocol: oops.

2006-06-20T17:22:56Z

Endy: STN Chemostat Protocol moved to Endy:STN Chemostat Protocol: oops.

←Older revision

Revision as of 17:22, 20 June 2006

Cconboy: S N Chemostat Protocol moved to Endy: STN Chemostat Protocol: name space

2006-06-20T17:19:06Z

S N Chemostat Protocol moved to Endy: STN Chemostat Protocol: name space

←Older revision

Revision as of 17:19, 20 June 2006

Cconboy at 22:12, 3 October 2005

2005-10-03T22:12:23Z

←Older revision		Revision as of 22:12, 3 October 2005
Line 1:		Line 1:
-	contact: [[Caitlin Conboy]], [[~~jen braff~~]]	+	contact: [[Caitlin Conboy]], [[Jennifer Braff]]
	==Day 0==		==Day 0==
	Set up o/n culture in supM9 media in a small flask at 37 C.		Set up o/n culture in supM9 media in a small flask at 37 C.

Cconboy at 22:11, 3 October 2005

2005-10-03T22:11:53Z

←Older revision		Revision as of 22:11, 3 October 2005
Line 1:		Line 1:
-	~~cmc_jcb~~	+	contact: [[Caitlin Conboy]], [[jen braff]]
	==Day 0==		==Day 0==
	Set up o/n culture in supM9 media in a small flask at 37 C.		Set up o/n culture in supM9 media in a small flask at 37 C.

Jcbraff at 16:10, 26 July 2005

2005-07-26T16:10:36Z

←Older revision		Revision as of 16:10, 26 July 2005
Line 6:		Line 6:
	==Day 1==		==Day 1==
	*Take OD600 of o/n culture. Dilute back in the morning into 25 mL/chamber supM9 (~1:100)		*Take OD600 of o/n culture. Dilute back in the morning into 25 mL/chamber supM9 (~1:100)
-	*When culture reaches ~OD 2.0 to add 20 mL to each chamber.	+	*When culture reaches ~OD 2.0 to add 20 mL to each chamber (usually late afternoon).
	*Run at 37 C. The bubbler should be set to the white dot for 6 chambers (med/high) and the effluent needle should be set to around the 24 mL mark to maintain a volume of 20 mL.		*Run at 37 C. The bubbler should be set to the white dot for 6 chambers (med/high) and the effluent needle should be set to around the 24 mL mark to maintain a volume of 20 mL.
	*Add the media input (teal, 23-G1 needle) and turn on pump (currently 10 rpm for 20 mL/chamber). After about 1 hour, check that the culture volume is 20 mL and adjust effluent as necessary.		*Add the media input (teal, 23-G1 needle) and turn on pump (currently 10 rpm for 20 mL/chamber). After about 1 hour, check that the culture volume is 20 mL and adjust effluent as necessary.
		+	*The cultures should wash out to a stable OD600 by the morning (day 2). OD600 should be monitored throughout the day (~every 3 hrs) and samples drawn for steady-state measurement either in the latter part of day 2 or on day 3.
		+

	==Notes==		==Notes==
Line 16:		Line 18:
	*Measure the flow rate by the effluent. 10 rpm gives about 0.25 mL/min, which is about right, but flow rate should be checked during each run.		*Measure the flow rate by the effluent. 10 rpm gives about 0.25 mL/min, which is about right, but flow rate should be checked during each run.
	*Mark 18 mL, 20 mL, and 22 mL on chemostat chambers to assist in monitoring volume.		*Mark 18 mL, 20 mL, and 22 mL on chemostat chambers to assist in monitoring volume.
-	*It is best to inoculate the chemostat in the early afternoon in order that OD600 may be monitored over the first several hours. The cultures should be at steady-state in the morning (day 2). OD600 should be monitored throughout the day (~every 2 hrs) and samples drawn for steady-state measurements in the afternoon.

Jcbraff at 16:02, 26 July 2005

2005-07-26T16:02:29Z

←Older revision		Revision as of 16:02, 26 July 2005
Line 1:		Line 1:
		+	cmc_jcb
	==Day 0==		==Day 0==
	Set up o/n culture in supM9 media in a small flask at 37 C.		Set up o/n culture in supM9 media in a small flask at 37 C.

Jcbraff at 16:00, 26 July 2005

2005-07-26T16:00:21Z

New page

==Day 0==
Set up o/n culture in supM9 media in a small flask at 37 C.

==Day 1==
*Take OD600 of o/n culture. Dilute back in the morning into 25 mL/chamber supM9 (~1:100)
*When culture reaches ~OD 2.0 to add 20 mL to each chamber.
*Run at 37 C. The bubbler should be set to the white dot for 6 chambers (med/high) and the effluent needle should be set to around the 24 mL mark to maintain a volume of 20 mL.
*Add the media input (teal, 23-G1 needle) and turn on pump (currently 10 rpm for 20 mL/chamber). After about 1 hour, check that the culture volume is 20 mL and adjust effluent as necessary.

==Notes==
*Run with bleach solution, then H20 after use. Autoclave all components on fast (22 psi, 250 F, 30 min.).
*This water bath should be filled with dI. Top up frequently with dI kept at 37 C.
*Use 1_5_05 supplemented M9 minimal media with AMP (50 ug/mL) or Kan (20 ug/mL) as needed.
*Measure the flow rate by the effluent. 10 rpm gives about 0.25 mL/min, which is about right, but flow rate should be checked during each run.
*Mark 18 mL, 20 mL, and 22 mL on chemostat chambers to assist in monitoring volume.
*It is best to inoculate the chemostat in the early afternoon in order that OD600 may be monitored over the first several hours. The cultures should be at steady-state in the morning (day 2). OD600 should be monitored throughout the day (~every 2 hrs) and samples drawn for steady-state measurements in the afternoon.