User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/09/25 - Revision history

Dhea Patel: /* Description */

Dhea Patel — Wed, 28 Nov 2012 19:19:55 GMT

Description

←Older revision		Revision as of 19:19, 28 November 2012
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	**25.0015g, 25.0013g, 24.9991g, and 24.9998g of LB were added to four separate Fernbach flasks.		**25.0015g, 25.0013g, 24.9991g, and 24.9998g of LB were added to four separate Fernbach flasks.
	**1.0L H<sub>2</sub>O was added to each flask.		**1.0L H<sub>2</sub>O was added to each flask.
-	**The flasks were covered with foil and labeled.	+	**The flasks were covered with foil and labeled A, B, C, and D, in order to distinguish each flask.
-		+
-	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 20:24, 7 October 2012 (EDT)''':how did you distigush and label the flasks? 1, 2, 3,4 or A, B, C, D?	+

	'''*[[AU_Biomaterials_Design_Lab:Protocols/Autoclave\|Autoclaving]]'''		'''*[[AU_Biomaterials_Design_Lab:Protocols/Autoclave\|Autoclaving]]'''
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	'''*Running Fluorescence'''		'''*Running Fluorescence'''
	*The fluorometer was set to have an excitation at 350nm (with a 15nm slit) and emission at 430nm (with a 20nm slit).		*The fluorometer was set to have an excitation at 350nm (with a 15nm slit) and emission at 430nm (with a 20nm slit).
		+	**The source was turned off because in chemiluminescence, the source is the sample itself.
	*The 13.2μL Phenol, 920μL Luminol, 990μL hydrogen peroxide, and 66μL HRP were added (the HRP was added at the last second) with the following concentrations:		*The 13.2μL Phenol, 920μL Luminol, 990μL hydrogen peroxide, and 66μL HRP were added (the HRP was added at the last second) with the following concentrations:
	[[Image:Screen_Shot_2012-09-26_at_2.42.56_PM.png]]		[[Image:Screen_Shot_2012-09-26_at_2.42.56_PM.png]]
-	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 20:24, 7 October 2012 (EDT)''':note that the source is turned off. becuase it is chemiluminescence the excitation comes form the chemical reaction not light.

	==Data==		==Data==

Dhea Patel: /* Data */

Dhea Patel — Thu, 25 Oct 2012 15:54:03 GMT

Data

←Older revision		Revision as of 15:54, 25 October 2012
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	**The HRP solution (23μM) was diluted to 2.3μM, 1.15μM, 0.23μM, and 23nM via serial dilution.		**The HRP solution (23μM) was diluted to 2.3μM, 1.15μM, 0.23μM, and 23nM via serial dilution.
	**The data obtained from the 2.3μM HRP:425μM Hydrogen Peroxide, 2.3μM HRP:1.7mM Hydrogen Peroxide, and 1.15μM HRP:1.7mM Hydrogen Peroxide (all containing 18mM phenol and 1.25mM Luminol) indicate that the reaction is occurring too quickly with the current HRP concentrations, as the slope of each of the lines created are too steep for effective analysis. The concentration of the HRP needs to be significantly decreased in order to obtain a clear curve.		**The data obtained from the 2.3μM HRP:425μM Hydrogen Peroxide, 2.3μM HRP:1.7mM Hydrogen Peroxide, and 1.15μM HRP:1.7mM Hydrogen Peroxide (all containing 18mM phenol and 1.25mM Luminol) indicate that the reaction is occurring too quickly with the current HRP concentrations, as the slope of each of the lines created are too steep for effective analysis. The concentration of the HRP needs to be significantly decreased in order to obtain a clear curve.
-
-	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 20:24, 7 October 2012 (EDT)''':data needs ot be in your notebook. you have nothing supporting your conclusions.

	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Dhea Patel: /* Data */

Dhea Patel — Thu, 25 Oct 2012 15:53:24 GMT

Data

←Older revision		Revision as of 15:53, 25 October 2012
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	*Please refer to [[User:Melissa_Novy/Notebook/CHEM-571/2012/09/25\| Melissa's entry]] for the results of the HRP Chemiluminescence Assay, including concentrations of each trial and a graph of intensity of phenol-enhanced solutions of luminol and hydrogen peroxide with HRP.		*Please refer to [[User:Melissa_Novy/Notebook/CHEM-571/2012/09/25\| Melissa's entry]] for the results of the HRP Chemiluminescence Assay, including concentrations of each trial and a graph of intensity of phenol-enhanced solutions of luminol and hydrogen peroxide with HRP.
		+
		+	[[Image:HRP_Assay_2012-09-25.png]]
		+
	**The HRP solution (23μM) was diluted to 2.3μM, 1.15μM, 0.23μM, and 23nM via serial dilution.		**The HRP solution (23μM) was diluted to 2.3μM, 1.15μM, 0.23μM, and 23nM via serial dilution.
	**The data obtained from the 2.3μM HRP:425μM Hydrogen Peroxide, 2.3μM HRP:1.7mM Hydrogen Peroxide, and 1.15μM HRP:1.7mM Hydrogen Peroxide (all containing 18mM phenol and 1.25mM Luminol) indicate that the reaction is occurring too quickly with the current HRP concentrations, as the slope of each of the lines created are too steep for effective analysis. The concentration of the HRP needs to be significantly decreased in order to obtain a clear curve.		**The data obtained from the 2.3μM HRP:425μM Hydrogen Peroxide, 2.3μM HRP:1.7mM Hydrogen Peroxide, and 1.15μM HRP:1.7mM Hydrogen Peroxide (all containing 18mM phenol and 1.25mM Luminol) indicate that the reaction is occurring too quickly with the current HRP concentrations, as the slope of each of the lines created are too steep for effective analysis. The concentration of the HRP needs to be significantly decreased in order to obtain a clear curve.

Abigail E. Miller at 00:24, 8 October 2012

Abigail E. Miller — Mon, 08 Oct 2012 00:24:59 GMT

←Older revision		Revision as of 00:24, 8 October 2012
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	**1.0L H<sub>2</sub>O was added to each flask.		**1.0L H<sub>2</sub>O was added to each flask.
	**The flasks were covered with foil and labeled.		**The flasks were covered with foil and labeled.
		+
		+	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 20:24, 7 October 2012 (EDT)''':how did you distigush and label the flasks? 1, 2, 3,4 or A, B, C, D?

	'''*[[AU_Biomaterials_Design_Lab:Protocols/Autoclave\|Autoclaving]]'''		'''*[[AU_Biomaterials_Design_Lab:Protocols/Autoclave\|Autoclaving]]'''
Line 65:		Line 67:
	*The 13.2μL Phenol, 920μL Luminol, 990μL hydrogen peroxide, and 66μL HRP were added (the HRP was added at the last second) with the following concentrations:		*The 13.2μL Phenol, 920μL Luminol, 990μL hydrogen peroxide, and 66μL HRP were added (the HRP was added at the last second) with the following concentrations:
	[[Image:Screen_Shot_2012-09-26_at_2.42.56_PM.png]]		[[Image:Screen_Shot_2012-09-26_at_2.42.56_PM.png]]
		+	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 20:24, 7 October 2012 (EDT)''':note that the source is turned off. becuase it is chemiluminescence the excitation comes form the chemical reaction not light.

	==Data==		==Data==
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	**The HRP solution (23μM) was diluted to 2.3μM, 1.15μM, 0.23μM, and 23nM via serial dilution.		**The HRP solution (23μM) was diluted to 2.3μM, 1.15μM, 0.23μM, and 23nM via serial dilution.
	**The data obtained from the 2.3μM HRP:425μM Hydrogen Peroxide, 2.3μM HRP:1.7mM Hydrogen Peroxide, and 1.15μM HRP:1.7mM Hydrogen Peroxide (all containing 18mM phenol and 1.25mM Luminol) indicate that the reaction is occurring too quickly with the current HRP concentrations, as the slope of each of the lines created are too steep for effective analysis. The concentration of the HRP needs to be significantly decreased in order to obtain a clear curve.		**The data obtained from the 2.3μM HRP:425μM Hydrogen Peroxide, 2.3μM HRP:1.7mM Hydrogen Peroxide, and 1.15μM HRP:1.7mM Hydrogen Peroxide (all containing 18mM phenol and 1.25mM Luminol) indicate that the reaction is occurring too quickly with the current HRP concentrations, as the slope of each of the lines created are too steep for effective analysis. The concentration of the HRP needs to be significantly decreased in order to obtain a clear curve.
		+
		+	*'''[[User:Abigail E. Miller\|Abigail E. Miller]] 20:24, 7 October 2012 (EDT)''':data needs ot be in your notebook. you have nothing supporting your conclusions.

	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Dhea Patel: /* Data */

Dhea Patel — Tue, 02 Oct 2012 18:42:26 GMT

Data

←Older revision		Revision as of 18:42, 2 October 2012
Line 69:		Line 69:

	*Please refer to [[User:Melissa_Novy/Notebook/CHEM-571/2012/09/25\| Melissa's entry]] for the results of the HRP Chemiluminescence Assay, including concentrations of each trial and a graph of intensity of phenol-enhanced solutions of luminol and hydrogen peroxide with HRP.		*Please refer to [[User:Melissa_Novy/Notebook/CHEM-571/2012/09/25\| Melissa's entry]] for the results of the HRP Chemiluminescence Assay, including concentrations of each trial and a graph of intensity of phenol-enhanced solutions of luminol and hydrogen peroxide with HRP.
-		+	**The HRP solution (23μM) was diluted to 2.3μM, 1.15μM, 0.23μM, and 23nM via serial dilution.
		+	**The data obtained from the 2.3μM HRP:425μM Hydrogen Peroxide, 2.3μM HRP:1.7mM Hydrogen Peroxide, and 1.15μM HRP:1.7mM Hydrogen Peroxide (all containing 18mM phenol and 1.25mM Luminol) indicate that the reaction is occurring too quickly with the current HRP concentrations, as the slope of each of the lines created are too steep for effective analysis. The concentration of the HRP needs to be significantly decreased in order to obtain a clear curve.

	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Dhea Patel: /* Description */

Dhea Patel — Wed, 26 Sep 2012 18:45:19 GMT

Description

←Older revision		Revision as of 18:45, 26 September 2012
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	*The fluorometer was set to have an excitation at 350nm (with a 15nm slit) and emission at 430nm (with a 20nm slit).		*The fluorometer was set to have an excitation at 350nm (with a 15nm slit) and emission at 430nm (with a 20nm slit).
	*The 13.2μL Phenol, 920μL Luminol, 990μL hydrogen peroxide, and 66μL HRP were added (the HRP was added at the last second) with the following concentrations:		*The 13.2μL Phenol, 920μL Luminol, 990μL hydrogen peroxide, and 66μL HRP were added (the HRP was added at the last second) with the following concentrations:
		+	[[Image:Screen_Shot_2012-09-26_at_2.42.56_PM.png]]

	==Data==		==Data==

Dhea Patel: /* Data */

Dhea Patel — Wed, 26 Sep 2012 18:35:16 GMT

Data

←Older revision		Revision as of 18:35, 26 September 2012
Line 67:		Line 67:
	==Data==		==Data==

-	Please refer to [[User:Melissa_Novy/Notebook/CHEM-571/2012/09/25\| Melissa's entry]] for the results of the HRP Chemiluminescence Assay, including concentrations of each trial and a graph of intensity of phenol-enhanced solutions of luminol and hydrogen peroxide with HRP.	+	*Please refer to [[User:Melissa_Novy/Notebook/CHEM-571/2012/09/25\| Melissa's entry]] for the results of the HRP Chemiluminescence Assay, including concentrations of each trial and a graph of intensity of phenol-enhanced solutions of luminol and hydrogen peroxide with HRP.

Dhea Patel: /* Data */

Dhea Patel — Wed, 26 Sep 2012 18:35:04 GMT

Data

←Older revision		Revision as of 18:35, 26 September 2012
Line 67:		Line 67:
	==Data==		==Data==

-	Please refer to [[User:Melissa_Novy/Notebook/CHEM-571/2012/09/25 Melissa's entry]] for the results of the HRP Chemiluminescence Assay, including concentrations of each trial and a graph of intensity of phenol-enhanced solutions of luminol and hydrogen peroxide with HRP.	+	Please refer to [[User:Melissa_Novy/Notebook/CHEM-571/2012/09/25\| Melissa's entry]] for the results of the HRP Chemiluminescence Assay, including concentrations of each trial and a graph of intensity of phenol-enhanced solutions of luminol and hydrogen peroxide with HRP.


	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
-	\|}	+

	__NOTOC__		__NOTOC__

Dhea Patel at 18:33, 26 September 2012

Dhea Patel — Wed, 26 Sep 2012 18:33:13 GMT

←Older revision		Revision as of 18:33, 26 September 2012
Line 65:		Line 65:
	*The 13.2μL Phenol, 920μL Luminol, 990μL hydrogen peroxide, and 66μL HRP were added (the HRP was added at the last second) with the following concentrations:		*The 13.2μL Phenol, 920μL Luminol, 990μL hydrogen peroxide, and 66μL HRP were added (the HRP was added at the last second) with the following concentrations:

		+	==Data==
		+
		+	Please refer to [[User:Melissa_Novy/Notebook/CHEM-571/2012/09/25 Melissa's entry]] for the results of the HRP Chemiluminescence Assay, including concentrations of each trial and a graph of intensity of phenol-enhanced solutions of luminol and hydrogen peroxide with HRP.

Dhea Patel: /* Description */

Dhea Patel — Tue, 25 Sep 2012 19:26:22 GMT

Description

←Older revision		Revision as of 19:26, 25 September 2012
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	*1mg of HRP was added to 1mL water, creating a 23μM HRP solution.		*1mg of HRP was added to 1mL water, creating a 23μM HRP solution.
	*The HRP was further diluted to 2.3μM by adding 1mL HRP solution to 9mL H<sub>2</sub>O.		*The HRP was further diluted to 2.3μM by adding 1mL HRP solution to 9mL H<sub>2</sub>O.
		+
		+	'''*Running Fluorescence'''
		+	*The fluorometer was set to have an excitation at 350nm (with a 15nm slit) and emission at 430nm (with a 20nm slit).
		+	*The 13.2μL Phenol, 920μL Luminol, 990μL hydrogen peroxide, and 66μL HRP were added (the HRP was added at the last second) with the following concentrations:
		+
		+

	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->