User:Elizabeth Ghias/Notebook/Experimental Biological Chemistry/2011/10/05

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Objective

To determine the fluorescence of MBP as a function of the concentration and to purify the DNA made from previously cultured cells.

Description

  1. 5 stock solutions, labeled A through E, were made by diluting MBP. The initial concentration of the MBP was determined to be 10.714 μM.
  2. Solution A was made by mixing 1397 μL of 10.714 μM MBP with 1603 μL of 50 mM Tris buffer (pH 7.55). The final concentration of solution A was 4.989 μM.
  3. Solution B was made by mixing 300 μL of solution A with 2700 μL of 50 mM Tris buffer (ph 7.55) for a final concentration of 0.4989 μM.
  4. Solution C was made by mixing 300 μL of solution B with 2700 μL of the same Tris buffer. The final concentration was 49.89 nM.
  5. Solution D was made by mixing 300 μL of solution C with 2700 μL of the Tris buffer for a concentration of 4.989 nM.
  6. Solution E was made by mixing 300 μL of solution D with 2700 μL of Tris buffer to give a final concentration of 0.4989 nM.
  7. The intensity of each solution was taken from 310 - 500 nm when excited at 290 nm. The intensity of a solution of 50 mM Tris buffer (ph 7.55) without MBP was also taken.

Data

Image:October_5_Corrected_Intensity_of_MBP_vs._Wavelength.png

The intensity values for each solution were corrected against the blank of 50 mM Tris (pH 7.55). The corrected intensities were graphed against the wavelengths. The intensity values of some of the high MBP concentrations were so high that they exceeded the maximum intensity that could be read by the fluorimeter.


The corrected intensities calculated for each MBP solution were then divided by the concentration of the solution. The corrected intensities divided by the concentrations were graphed against the wavelengths for each solution. Image:Solution_A_Corrected_Intensity_Divided_by_Concentration_vs._Wavelength.png

Image:Solution_B_Corrected_Intensity_Divided_by_Wavelength.png

Image:Solution_C_Corrected_Intensity_Divided_by_Concentration_vs._Wavelength.png

Image:Solution_D_Corrected_Intensity_Divided_by_Concentration_vs._Wavelength.png

Image:Solution_E_Corrected_Intensity_Divided_by_Concentration_vs._Wavelength.png


Notes

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