User:Floriane Briere/Notebook/CHEM-496/2012/03/06

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Objective

Today's objective is to perform the first part of the tagging reaction which will be finished tomorrow so that the dialysis will be performed during the spring break week. So, we are going to use the 3 same solutions (70 ratio, 166 ratio and BSA/HCl control) and labelled them with the same dye. Then, after the spring break will perform fluorescence spectra to check if our results can be reproduced or enhanced.

Protocol

We are going to use solutions prepared on the 8th of february.

  • Preparing 3 dye solutions: 2.0mg of dye + 160µL of DMSO
  • Changing the pH of solutions:
  1. 70 ratio solution:
    1. Initial: pH = 3.75 and volume = 10mL
    2. Added 600µL of Tris buffer (pH = 8.50)
    3. Final: pH = 8.45 and volume = 10.6mL
  2. 166 ratio solution:
    1. Initial: pH = 3.15 and volume = 10mL
    2. Added 1.5mL of Tris buffer (pH = 8.50)
    3. Final: pH = 8.44 and volume = 11.5mL
  3. BSA control (BSA/HCl solution):
    1. Initial: pH = 4.37 and volume = 10mL
    2. Added 300µL of Tris buffer (pH = 8.50)
    3. Final: pH = 8.41 and volume = 10.3mL
  • Dying reaction:
  1. Add to each solution 160µL of the dye solution
  2. Incubation at room temperature for 24 hours
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