User:Elizabeth Ghias/Notebook/Experimental Biological Chemistry/2012/03/27: Difference between revisions

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==Objective==
==Objective==
To adjust the pHs of the solutions so that they are all equal and to retake UV-Vis and fluorescence spectra
To adjust the pHs of all the dye solutions (both first and second trial) so that they are all equal and to retake UV-Vis and fluorescence spectra


==Description==
==Description==
# Add 50 mM tris buffer (pH = 8.73) to raise the pHs of the samples to a pH around 8.30.
1. The initial pHs of the solutions from both the first and second trials were measured.
# The samples containing fibers (166 molar ratio) were centrifuged for 5 minutes at 13,200 rpg.
 
# Fluorescence spectra were taken of the samples between 620 - 800 nm, with excitation at 600 nm.
<u>First Trial Initial pHs</u>
* 70 molar ratio with dye pH = 8.12
* 166 molar ratio with dye pH = 8.43
* BSA/HCl with dye pH 8.30
 
<u>Second Trial Initial pHs</u>
* 70 molar ratio with dye pH = 8.00
* 166 molar ratio with dye pH = 8.13
* BSA/HCl with dye pH = 7.98
 
2. Add 50 mM tris buffer (pH = 8.73) to raise the pHs of the samples to a pH around 8.30.
 
<u>First Trial pH Changes</u>
* 70 molar ratio with dye: 10 drops of Tris were added to give a final pH of 8.30
 
<u>Second Trial pH Changes</u>
* 70 molar ratio with dye: 6 drops of Tris added, final pH = 8.29
* 166 molar ratio with dye: 3 drops of Tris added, final pH = 8.41
* BSA/HCl with dye: 7 drops of Tris added, final pH = 8.28
 
3. The samples containing fibers (all dye samples) were centrifuged for 5 minutes at 13,200 rpg.
 
4. Fluorescence spectra were taken of the samples between 620 - 800 nm, with excitation at 600 nm.
 
5. Fluorescence spectra were also taken of the 70 molar ratio, 166 molar ratio, and the BSA/HCl reactions without dye.
 
6. UV-Vis spectra were taken of all dye and control solutions.




==Data==
==Data==
* Add data and results here...
<u>Fluorescence Spectra of Solutions from the Second Trial</u>
 
[[Image:27mar - fluorescence curve 2nd solution.jpg]]


==Notes==
* When the pHs were initially measured, the values had changed from the week before
* The fluorescence spectra of the 70 molar ratio is now slightly higher than that of the 166 molar ratio


<u>Fluorescence Spectra of Solutions from the Initial Trial</u>
[[Image:27mar - fluorescence curve initial solution.jpg]]
<u>Fluorescence Spectrum of Control solutions</u>
[[Image:27mar - fluorescence curve control solution.jpg]]
<u>UV Spectra of Solutions from the Second Trial</u>
[[Image:27mar - UV curve second solution.jpg]]
[[Image:27mar - UV curve second solution zoom.jpg]]
<u>UV Spectra of Solutions from the Initial Trial</u>
[[Image:27mar - UV curve initial solution.jpg]]
[[Image:27mar - UV curve initial solution zoom.jpg]]
<u>UV Spectrum of Control Solutions</u>
[[Image:27mar - UV curve control solution.jpg]]


Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.
[[Image:27mar - UV curve control solution zoom.jpg]]


[[Category:Course]]
==Notes==
[[Category:Miscellaneous]]
* When the pHs were initially measured, the values had changed from the week before. For example, the 166 molar ratio solution with dye from the second trial had a measured pH of 8.35 the previous week, but this week had a decreased pH of 8.13
* The changes in pH values could be explained by contamination or a reaction that is occurring in the dye solutions.
* The fluorescence spectra of the 70 molar ratio with dye now has a slightly higher intensity than that of the 166 molar ratio with dye.  The BSA/HCl dye mixture now has the lowest intensity




Revision as of 11:04, 10 April 2012

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Objective

To adjust the pHs of all the dye solutions (both first and second trial) so that they are all equal and to retake UV-Vis and fluorescence spectra

Description

1. The initial pHs of the solutions from both the first and second trials were measured.

First Trial Initial pHs

  • 70 molar ratio with dye pH = 8.12
  • 166 molar ratio with dye pH = 8.43
  • BSA/HCl with dye pH 8.30

Second Trial Initial pHs

  • 70 molar ratio with dye pH = 8.00
  • 166 molar ratio with dye pH = 8.13
  • BSA/HCl with dye pH = 7.98

2. Add 50 mM tris buffer (pH = 8.73) to raise the pHs of the samples to a pH around 8.30.

First Trial pH Changes

  • 70 molar ratio with dye: 10 drops of Tris were added to give a final pH of 8.30

Second Trial pH Changes

  • 70 molar ratio with dye: 6 drops of Tris added, final pH = 8.29
  • 166 molar ratio with dye: 3 drops of Tris added, final pH = 8.41
  • BSA/HCl with dye: 7 drops of Tris added, final pH = 8.28

3. The samples containing fibers (all dye samples) were centrifuged for 5 minutes at 13,200 rpg.

4. Fluorescence spectra were taken of the samples between 620 - 800 nm, with excitation at 600 nm.

5. Fluorescence spectra were also taken of the 70 molar ratio, 166 molar ratio, and the BSA/HCl reactions without dye.

6. UV-Vis spectra were taken of all dye and control solutions.


Data

Fluorescence Spectra of Solutions from the Second Trial


Fluorescence Spectra of Solutions from the Initial Trial


Fluorescence Spectrum of Control solutions


UV Spectra of Solutions from the Second Trial


UV Spectra of Solutions from the Initial Trial


UV Spectrum of Control Solutions

Notes

  • When the pHs were initially measured, the values had changed from the week before. For example, the 166 molar ratio solution with dye from the second trial had a measured pH of 8.35 the previous week, but this week had a decreased pH of 8.13
  • The changes in pH values could be explained by contamination or a reaction that is occurring in the dye solutions.
  • The fluorescence spectra of the 70 molar ratio with dye now has a slightly higher intensity than that of the 166 molar ratio with dye. The BSA/HCl dye mixture now has the lowest intensity



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