User:Michael F. Nagle/Notebook/Chem 571/2012/09/18 - Revision history

Michael F. Nagle at 00:44, 8 December 2012

2012-12-08T00:44:44Z

←Older revision		Revision as of 00:44, 8 December 2012
Line 24:		Line 24:
	##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer.		##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer.
	#The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.		#The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.
-	#E. Coli with a plasmid coding for Adenosine Deaminase (ADA)was inserted into the broth.	+	#E. Coli transformed with a plasmid coding for Adenosine Deaminase (ADA)was inserted into the broth.
	#The flasks were left overnight in an orbital incubator shaker at 37°C and 237 rpm		#The flasks were left overnight in an orbital incubator shaker at 37°C and 237 rpm

Michael F. Nagle: /* Procedure */

2012-10-26T03:05:46Z

Procedure

←Older revision		Revision as of 03:05, 26 October 2012
Line 17:		Line 17:
	#Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them.		#Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them.
	#Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them.		#Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them.
-	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 11:25, 7 October 2012 (EDT)''':inidiacte how you distigushedthe flasks..1, 2, 3, 4? or A, B, C, D? how do you know if htey act differently due to the amounts of chemicals added?
	#Amount of Kanamycin for cultures was calculated. Kanamycin is an antibiotic and is being used to prevent microbial growth besides that of E. Coli.		#Amount of Kanamycin for cultures was calculated. Kanamycin is an antibiotic and is being used to prevent microbial growth besides that of E. Coli.
	##The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth.		##The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth.
Line 25:		Line 24:
	##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer.		##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer.
	#The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.		#The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.
-	#~~The~~ plasmid ~~that codes~~ for Adenosine Deaminase (ADA)was inserted into the broth.	+	#E. Coli with a plasmid coding for Adenosine Deaminase (ADA)was inserted into the broth.
-	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 11:25, 7 October 2012 (EDT)''':plasmid is in the e. coli, therefore you add a colony of E. coli cells that contain the plasmid for ADA. split?	+	#The flasks were left overnight in an orbital incubator shaker at 37°C and 237 rpm
-	#The flasks were ~~placed~~ in an orbital incubator shaker at 37°C and 237 rpm	+
-	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 11:25, 7 October 2012 (EDT)''':not that this was left to shake overnight	+

	==Discussion==		==Discussion==

Abigail E. Miller: /* Procedure */

2012-10-07T15:25:11Z

Procedure

←Older revision		Revision as of 15:25, 7 October 2012
Line 17:		Line 17:
	#Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them.		#Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them.
	#Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them.		#Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them.
		+	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 11:25, 7 October 2012 (EDT)''':inidiacte how you distigushedthe flasks..1, 2, 3, 4? or A, B, C, D? how do you know if htey act differently due to the amounts of chemicals added?
	#Amount of Kanamycin for cultures was calculated. Kanamycin is an antibiotic and is being used to prevent microbial growth besides that of E. Coli.		#Amount of Kanamycin for cultures was calculated. Kanamycin is an antibiotic and is being used to prevent microbial growth besides that of E. Coli.
	##The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth.		##The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth.
Line 25:		Line 26:
	#The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.		#The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.
	#The plasmid that codes for Adenosine Deaminase (ADA)was inserted into the broth.		#The plasmid that codes for Adenosine Deaminase (ADA)was inserted into the broth.
		+	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 11:25, 7 October 2012 (EDT)''':plasmid is in the e. coli, therefore you add a colony of E. coli cells that contain the plasmid for ADA. split?
	#The flasks were placed in an orbital incubator shaker at 37°C and 237 rpm		#The flasks were placed in an orbital incubator shaker at 37°C and 237 rpm
		+	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 11:25, 7 October 2012 (EDT)''':not that this was left to shake overnight

	==Discussion==		==Discussion==

Michael F. Nagle: /* Procedure */

2012-10-06T21:45:21Z

Procedure

Michael F. Nagle at 04:11, 5 October 2012

2012-10-05T04:11:17Z

←Older revision		Revision as of 04:11, 5 October 2012
Line 25:		Line 25:
	#The plasmid that codes for Adenosine Deaminase (ADA)was inserted into the broth.		#The plasmid that codes for Adenosine Deaminase (ADA)was inserted into the broth.
	#The flasks were placed in an orbital incubator shaker at 37°C and 237 rpm		#The flasks were placed in an orbital incubator shaker at 37°C and 237 rpm
-		+	==Discussion==
		+	*ADA is being synthesised for upcoming experiments on it's potential ability to make gold into nanoparticles.

Michael F. Nagle: /* Procedure */

2012-10-01T23:38:37Z

Procedure

Michael F. Nagle at 23:34, 1 October 2012

2012-10-01T23:34:07Z

←Older revision		Revision as of 23:34, 1 October 2012
Line 10:		Line 10:
	==Procedure==		==Procedure==
	#Starter Culture and Expression Culture		#Starter Culture and Expression Culture
-	##[http://openwetware.org/wiki/AU_Biomaterials_Design_Lab:Protocols/Starter_Culture_Media ~~starter~~ culture media]and [http://openwetware.org/wiki/AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media expression culture media] were made following procedure. 25mL water was used instead of 35mL in making the starter culture.	+	##[http://openwetware.org/wiki/AU_Biomaterials_Design_Lab:Protocols/Starter_Culture_Media Starter culture media]and [http://openwetware.org/wiki/AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media expression culture media] were made following procedure. 25mL water was used instead of 35mL in making the starter culture.
	##The broth flasks were capped with foil.		##The broth flasks were capped with foil.
	##The flasks were put in the autoclave at 121°C for 60 minutes. Due to limited space, the flasks had to be autoclaved in two batches.		##The flasks were put in the autoclave at 121°C for 60 minutes. Due to limited space, the flasks had to be autoclaved in two batches.

Michael F. Nagle at 18:37, 1 October 2012

2012-10-01T18:37:04Z

←Older revision		Revision as of 18:37, 1 October 2012
Line 9:		Line 9:
	* Grow E. Coli in [[AU Biomaterials Design Lab:Materials/LB\|LB]] broth, for the purpose of expressing ADA.		* Grow E. Coli in [[AU Biomaterials Design Lab:Materials/LB\|LB]] broth, for the purpose of expressing ADA.
	==Procedure==		==Procedure==
-	# ~~In four 2.8L flasks, broth was created with 25g [~~[~~AU Biomaterials Design Lab~~:~~Materials~~/~~LB\|LB]~~] and ~~1L H<sub>2</sub>O, using the protocol to make [~~[AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media\|expression culture]]. ~~Four more smaller 250mL~~ flasks were ~~prepared using this protocol~~ for ~~making [[AU_Biomaterials_Design_Lab:Protocols/Starter_Culture_Media\|Starter Culture]]~~	+	#Starter Culture and Expression Culture
-	# The ~~starter culture~~ was ~~steamed~~ in the autoclave ~~following [[AU Biomaterials Design Lab:Protocols~~/~~Autoclave\|this protocol]]~~	+	##[http://openwetware.org/wiki/AU_Biomaterials_Design_Lab:Protocols/Starter_Culture_Media starter culture media]and [http://openwetware.org/wiki/AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media expression culture media] were made following procedure. 25mL water was used instead of 35mL in making the starter culture.
		+	##The broth flasks were capped with foil.
		+	##The flasks were put in the autoclave at 121°C for 60 minutes. Due to limited space, the flasks had to be autoclaved in two batches.
		+	##The second round only went in for 50 minutes because the autoclave was still warm. After the autoclaving was complete for each round, the flasks were left in for 30 minutes while the autoclave depressurized. The pressure meter was confirmed to be at 0 before the door was opened.
		+	##Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them.
		+	##Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them.
		+	##Amount of Kanamycin for cultures was calculated. The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth.
		+	##50mg/mL Kanamycin * 1µg/.001mg = 50,000µg/mL Kanamycin
		+	##50µg/mL * 35mL = 50000µg/mL
		+	##.035mL Kanamycin
		+	##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer.
		+	##The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.
		+	##The plasmid that codes for Adenosine Deaminase (ADA)was inserted into the broth.
		+	##The flasks were placed in an orbital incubator shaker at 37°C and 237 rpm
		+

Michael F. Nagle: /* Procedure */

2012-09-19T17:39:10Z

Procedure

←Older revision		Revision as of 17:39, 19 September 2012
Line 9:		Line 9:
	* Grow E. Coli in [[AU Biomaterials Design Lab:Materials/LB\|LB]] broth, for the purpose of expressing ADA.		* Grow E. Coli in [[AU Biomaterials Design Lab:Materials/LB\|LB]] broth, for the purpose of expressing ADA.
	==Procedure==		==Procedure==
-	# In four 2.8L flasks, broth was created with 25g [[AU Biomaterials Design Lab:Materials/LB\|LB]] and 1L H<sub>2</sub>O. Four more smaller 250mL flasks were prepared using this protocol for making [[AU_Biomaterials_Design_Lab:Protocols/Starter_Culture_Media\|Starter Culture]]	+	# In four 2.8L flasks, broth was created with 25g [[AU Biomaterials Design Lab:Materials/LB\|LB]] and 1L H<sub>2</sub>O, using the protocol to make [[AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media\|expression culture]]. Four more smaller 250mL flasks were prepared using this protocol for making [[AU_Biomaterials_Design_Lab:Protocols/Starter_Culture_Media\|Starter Culture]]
	# The starter culture was steamed in the autoclave following [[AU Biomaterials Design Lab:Protocols/Autoclave\|this protocol]]		# The starter culture was steamed in the autoclave following [[AU Biomaterials Design Lab:Protocols/Autoclave\|this protocol]]

Michael F. Nagle at 18:10, 18 September 2012

2012-09-18T18:10:21Z

←Older revision		Revision as of 18:10, 18 September 2012
Line 6:		Line 6:
	\| 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~~==	+	==Objectives==
-	* ~~Insert content here~~...	+	* Grow E. Coli in [[AU Biomaterials Design Lab:Materials/LB\|LB]] broth, for the purpose of expressing ADA.
		+	==Procedure==
		+	# In four 2.8L flasks, broth was created with 25g [[AU Biomaterials Design Lab:Materials/LB\|LB]] and 1L H<sub>2</sub>O. Four more smaller 250mL flasks were prepared using this protocol for making [[AU_Biomaterials_Design_Lab:Protocols/Starter_Culture_Media\|Starter Culture]]
		+	# The starter culture was steamed in the autoclave following [[AU Biomaterials Design Lab:Protocols/Autoclave\|this protocol]]

←Older revision		Revision as of 21:45, 6 October 2012
Line 17:		Line 17:
	#Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them.		#Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them.
	#Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them.		#Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them.
-	#Amount of Kanamycin for cultures was calculated. The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth.	+	#Amount of Kanamycin for cultures was calculated. Kanamycin is an antibiotic and is being used to prevent microbial growth besides that of E. Coli.
		+	##The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth.
	##50mg/mL Kanamycin * 1µg/.001mg = 50,000µg/mL Kanamycin		##50mg/mL Kanamycin * 1µg/.001mg = 50,000µg/mL Kanamycin
-	##50µg/mL * 35mL = 50000µg/mL	+	##50µg/mL * 35mL = 50000µg/mL * volume needed
-	##.035mL Kanamycin	+	##.035mL Kanamycin solution needed
	##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer.		##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer.
	#The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.		#The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.
	#The plasmid that codes for Adenosine Deaminase (ADA)was inserted into the broth.		#The plasmid that codes for Adenosine Deaminase (ADA)was inserted into the broth.
-	#The flasks were placed in an orbital incubator shaker at 37°C and 237 rpm	+	#The flasks were placed in an orbital incubator shaker at 37°C and 237 rpm
		+
	==Discussion==		==Discussion==
	*ADA is being synthesised for upcoming experiments on it's potential ability to make gold into nanoparticles.		*ADA is being synthesised for upcoming experiments on it's potential ability to make gold into nanoparticles.

←Older revision		Revision as of 23:38, 1 October 2012
Line 9:		Line 9:
	* Grow E. Coli in [[AU Biomaterials Design Lab:Materials/LB\|LB]] broth, for the purpose of expressing ADA.		* Grow E. Coli in [[AU Biomaterials Design Lab:Materials/LB\|LB]] broth, for the purpose of expressing ADA.
	==Procedure==		==Procedure==
-	#Starter Culture and Expression Culture	+	Starter Culture and Expression Culture
-	##[http://openwetware.org/wiki/AU_Biomaterials_Design_Lab:Protocols/Starter_Culture_Media Starter culture media]and [http://openwetware.org/wiki/AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media expression culture media] were made following procedure. 25mL water was used instead of 35mL in making the starter culture.	+	#[http://openwetware.org/wiki/AU_Biomaterials_Design_Lab:Protocols/Starter_Culture_Media Starter culture media]and [http://openwetware.org/wiki/AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media expression culture media] were made following procedure. 25mL water was used instead of 35mL in making the starter culture.
-	##The broth flasks were capped with foil.	+	#The broth flasks were capped with foil.
-	##The flasks were put in ~~the autoclave~~ at 121°C for 60 minutes. Due to limited space, the flasks had to be autoclaved in two batches.	+	#Autoclave
		+	##The flasks were put in at 121°C for 60 minutes. Due to limited space, the flasks had to be autoclaved in two batches.
	##The second round only went in for 50 minutes because the autoclave was still warm. After the autoclaving was complete for each round, the flasks were left in for 30 minutes while the autoclave depressurized. The pressure meter was confirmed to be at 0 before the door was opened.		##The second round only went in for 50 minutes because the autoclave was still warm. After the autoclaving was complete for each round, the flasks were left in for 30 minutes while the autoclave depressurized. The pressure meter was confirmed to be at 0 before the door was opened.
-	##Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them.	+	#Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them.
-	##Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them.	+	#Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them.
-	##Amount of Kanamycin for cultures was calculated. The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth.	+	#Amount of Kanamycin for cultures was calculated. The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth.
	##50mg/mL Kanamycin * 1µg/.001mg = 50,000µg/mL Kanamycin		##50mg/mL Kanamycin * 1µg/.001mg = 50,000µg/mL Kanamycin
	##50µg/mL * 35mL = 50000µg/mL		##50µg/mL * 35mL = 50000µg/mL
	##.035mL Kanamycin		##.035mL Kanamycin
	##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer.		##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer.
-	##The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.	+	#The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time.
-	##The plasmid that codes for Adenosine Deaminase (ADA)was inserted into the broth.	+	#The plasmid that codes for Adenosine Deaminase (ADA)was inserted into the broth.
-	##The flasks were placed in an orbital incubator shaker at 37°C and 237 rpm	+	#The flasks were placed in an orbital incubator shaker at 37°C and 237 rpm