User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/16 - Revision history

Mary Mendoza: /* Continuation of Chemiluminescence */

Mary Mendoza — Fri, 07 Dec 2012 14:00:06 GMT

Continuation of Chemiluminescence

←Older revision		Revision as of 14:00, 7 December 2012
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	{\| {{table}}		{\| {{table}}
		+	\| align="center" style="background:#f0f0f0;"\|''''
	\| align="center" style="background:#f0f0f0;"\|''HRP''		\| align="center" style="background:#f0f0f0;"\|''HRP''
	\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''		\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''
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	\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''		\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''
	\|-		\|-
-	\| 33 μL\|\|27 μL\|\|450 μL\|\|88 μL\|\|902 μL	+	\| Molarity of stock solutions\|\|2.3 μM\|\|18 mM\|\|10 mM\|\|1.7 mM\|\|neutral
		+	\|-
		+	\| Volume\|\|33 μL\|\|27 μL\|\|450 μL\|\|88 μL\|\|902 μL
	\|-		\|-

Mary Mendoza: /* PCR mutation */

Mary Mendoza — Sat, 27 Oct 2012 15:11:06 GMT

PCR mutation

←Older revision		Revision as of 15:11, 27 October 2012
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-	* The procedure listed in [[AU Biomaterials Design Lab:Protocols/PCR Mutation Protocol\|PCR Mutation protocol]] was strictly followed~~. The amplified DNA was contained in a sterilized, 1.5 microcentrifuge tube~~. After the addition of all reagents, the sample was placed in the thermocycler.	+	* The procedure listed in [[AU Biomaterials Design Lab:Protocols/PCR Mutation Protocol\|PCR Mutation protocol]] was strictly followed. After the addition of all reagents, the sample was placed in the thermocycler. The amplified DNA was contained in a sterilized, 1.5 microcentrifuge tube.

	==Continuation of Chemiluminescence==		==Continuation of Chemiluminescence==

Mary Mendoza: /* PCR mutation */

Mary Mendoza — Sat, 27 Oct 2012 15:09:22 GMT

PCR mutation

←Older revision		Revision as of 15:09, 27 October 2012
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-	* The procedure listed in [[AU Biomaterials Design Lab:Protocols/PCR Mutation Protocol\|PCR Mutation protocol]] was strictly followed. After the addition of all reagents, the sample was placed in the thermocycler.	+	* The procedure listed in [[AU Biomaterials Design Lab:Protocols/PCR Mutation Protocol\|PCR Mutation protocol]] was strictly followed. The amplified DNA was contained in a sterilized, 1.5 microcentrifuge tube. After the addition of all reagents, the sample was placed in the thermocycler.

	==Continuation of Chemiluminescence==		==Continuation of Chemiluminescence==

Mary Mendoza: /* PCR mutation */

Mary Mendoza — Sat, 27 Oct 2012 04:18:24 GMT

PCR mutation

←Older revision		Revision as of 04:18, 27 October 2012
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-	<math>\frac{0.46E6 ng}{x \mu L}</math> = <math>\frac{100 ng}{1 \mu L}</math> of primer in water = 4600 μL of water	+	<math>\frac{0.46E6 ng}{x \mu L}</math> = <math>\frac{100 ng}{1 \mu L}</math> of primer in water = 4600 μL of water

Mary Mendoza: /* PCR mutation */

Mary Mendoza — Fri, 26 Oct 2012 16:55:08 GMT

PCR mutation

←Older revision		Revision as of 16:55, 26 October 2012
Line 22:		Line 22:


-	* The procedure listed in [[AU Biomaterials Design Lab:Protocols/PCR Mutation Protocol\|PCR Mutation protocol]] was strictly followed. After the addition of all reagents, the sample was placed in the thermocycler.	+	* The procedure listed in [[AU Biomaterials Design Lab:Protocols/PCR Mutation Protocol\|PCR Mutation protocol]] was strictly followed. After the addition of all reagents, the sample was placed in the thermocycler.

	==Continuation of Chemiluminescence==		==Continuation of Chemiluminescence==

Mary Mendoza: /* Continuation of Chemiluminescence */

Mary Mendoza — Fri, 26 Oct 2012 07:01:32 GMT

Continuation of Chemiluminescence

←Older revision		Revision as of 07:01, 26 October 2012
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	* Due to time constraints, the chemiluminescence of the luminol at pH 10.55 was tested on a lab bench with the room lights turned off. The reaction produced a neon blue glow that lasted for more than 3 minutes. No photograph was taken since the apparatus of the camera was not suited for the fluorescence activity.		* Due to time constraints, the chemiluminescence of the luminol at pH 10.55 was tested on a lab bench with the room lights turned off. The reaction produced a neon blue glow that lasted for more than 3 minutes. No photograph was taken since the apparatus of the camera was not suited for the fluorescence activity.
		+
		+	* The volume of each reagent added to the cuvette are listed below.

	{\| {{table}}		{\| {{table}}

Mary Mendoza: /* Continuation of Chemiluminescence */

Mary Mendoza — Fri, 26 Oct 2012 06:58:22 GMT

Continuation of Chemiluminescence

←Older revision		Revision as of 06:58, 26 October 2012
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	* Due to time constraints, the chemiluminescence of the luminol at pH 10.55 was tested on a lab bench with the room lights turned off. The reaction produced a neon blue glow that lasted for more than 3 minutes. No photograph was taken since the apparatus of the camera was not suited for the fluorescence activity.		* Due to time constraints, the chemiluminescence of the luminol at pH 10.55 was tested on a lab bench with the room lights turned off. The reaction produced a neon blue glow that lasted for more than 3 minutes. No photograph was taken since the apparatus of the camera was not suited for the fluorescence activity.

-		+	{\| {{table}}
		+	\| align="center" style="background:#f0f0f0;"\|''HRP''
		+	\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''
		+	\| align="center" style="background:#f0f0f0;"\|''luminol''
		+	\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O<sub>2</sub>''
		+	\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''
		+	\|-
		+	\| 33 μL\|\|27 μL\|\|450 μL\|\|88 μL\|\|902 μL
		+	\|-

Mary Mendoza: /* Continuation of Chemiluminescence */

Mary Mendoza — Fri, 26 Oct 2012 06:54:39 GMT

Continuation of Chemiluminescence

←Older revision		Revision as of 06:54, 26 October 2012
Line 43:		Line 43:

	* Due to time constraints, the chemiluminescence of the luminol at pH 10.55 was tested on a lab bench with the room lights turned off. The reaction produced a neon blue glow that lasted for more than 3 minutes. No photograph was taken since the apparatus of the camera was not suited for the fluorescence activity.		* Due to time constraints, the chemiluminescence of the luminol at pH 10.55 was tested on a lab bench with the room lights turned off. The reaction produced a neon blue glow that lasted for more than 3 minutes. No photograph was taken since the apparatus of the camera was not suited for the fluorescence activity.
		+
		+

Mary Mendoza: /* Continuation of Chemiluminescence */

Mary Mendoza — Fri, 26 Oct 2012 06:53:13 GMT

Continuation of Chemiluminescence

←Older revision		Revision as of 06:53, 26 October 2012
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	* A weight of .0112 g of luminol was added to 6 mL of water. The buffer composed of a direct addition of .0737 g of sodium carbonate and .4358 g of sodium bicarbonate.		* A weight of .0112 g of luminol was added to 6 mL of water. The buffer composed of a direct addition of .0737 g of sodium carbonate and .4358 g of sodium bicarbonate.
	* Using a pH meter, the electrode detected the pH at 8.73. As suggested by Dr. Hartings, a solution of sodium carbonate was made to increase the pH of the solution.		* Using a pH meter, the electrode detected the pH at 8.73. As suggested by Dr. Hartings, a solution of sodium carbonate was made to increase the pH of the solution.
-	* Several adjustments were made in increasing the pH. A total weight of 1.91 g of sodium carbonate dissolved in 15 mL of water was added to the 6 mL solution of luminol.	+	* Several adjustments were made in increasing the pH. A total weight of 1.91 g of sodium carbonate dissolved in 15 mL of water was added to the 6 mL solution of luminol. The final pH for luminol was 10.55.
	* The molarity of sodium carbonate (MW 105.9784 g/mol) added was calculated:		* The molarity of sodium carbonate (MW 105.9784 g/mol) added was calculated:

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-	* Since there was limited amounts of the solid form of luminol, it was decided to take 6 mL of the 2.86 mM luminol stock solution and then add the appropriate amount of luminol to the 6 mL volume. The molarity of the 6 mL solution was very minute; the molarity was approximated as 2 mM. By making this assumption, .00106 g of luminol would make a 1 mM solution in 6 mL of water; this amount was multiplied by 8. The product was 0.00848 g of luminol was needed to be added into the 2 mM solution of luminol to increase the molarity to 10 mM.	+	* Since there was limited amounts of the solid form of luminol, it was decided to take 6 mL of the 2.86 mM luminol stock solution and then add the appropriate amount of luminol to the 6 mL volume. The molarity of the 6 mL solution was very minute; the molarity was approximated as 2 mM. By making this assumption, .00106 g of luminol would make a 1 mM solution in 6 mL of water; this amount was multiplied by 8. The product was 0.00848 g of luminol was needed to be added into the 2 mM solution of luminol to increase the molarity to 10 mM. The amount was weighed and added to the 6 mL 2.86 mM solution of luminol.
		+
		+	* Due to time constraints, the chemiluminescence of the luminol at pH 10.55 was tested on a lab bench with the room lights turned off. The reaction produced a neon blue glow that lasted for more than 3 minutes. No photograph was taken since the apparatus of the camera was not suited for the fluorescence activity.

Mary Mendoza: /* Continuation of Chemiluminescence */

Mary Mendoza — Fri, 26 Oct 2012 06:48:54 GMT

Continuation of Chemiluminescence

←Older revision		Revision as of 06:48, 26 October 2012
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	* The concentration of luminol has been diluted by the addition of 15 mL of water.		* The concentration of luminol has been diluted by the addition of 15 mL of water.
		+

	Molarity of diluted luminol = <math>\frac{10 mM * 6 mL}{21 mL}</math> = 2.85714 mM of luminol		Molarity of diluted luminol = <math>\frac{10 mM * 6 mL}{21 mL}</math> = 2.85714 mM of luminol
		+
		+
		+	* Since there was limited amounts of the solid form of luminol, it was decided to take 6 mL of the 2.86 mM luminol stock solution and then add the appropriate amount of luminol to the 6 mL volume. The molarity of the 6 mL solution was very minute; the molarity was approximated as 2 mM. By making this assumption, .00106 g of luminol would make a 1 mM solution in 6 mL of water; this amount was multiplied by 8. The product was 0.00848 g of luminol was needed to be added into the 2 mM solution of luminol to increase the molarity to 10 mM.
		+
		+

←Older revision		Revision as of 14:00, 7 December 2012
Line 47:		Line 47:

	{\| {{table}}		{\| {{table}}
		+	\| align="center" style="background:#f0f0f0;"\|''''
	\| align="center" style="background:#f0f0f0;"\|''HRP''		\| align="center" style="background:#f0f0f0;"\|''HRP''
	\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''		\| align="center" style="background:#f0f0f0;"\|''4-iodophenol''
Line 53:		Line 54:
	\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''		\| align="center" style="background:#f0f0f0;"\|''H<sub>2</sub>O''
	\|-		\|-
-	\| 33 μL\|\|27 μL\|\|450 μL\|\|88 μL\|\|902 μL	+	\| Molarity of stock solutions\|\|2.3 μM\|\|18 mM\|\|10 mM\|\|1.7 mM\|\|neutral
		+	\|-
		+	\| Volume\|\|33 μL\|\|27 μL\|\|450 μL\|\|88 μL\|\|902 μL
	\|-		\|-