User:Floriane Briere/Notebook/CHEM-496/2012/02/15
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ObjectiveToday's objective is to perform a dialysis using the solution we made yesterday. The reaction has been running for 24hours in the shaker, allowing the dye to tag our BSA peptides by binding unprotonated lysine. The dialysis is going to allow us to remove the excess of dye which hasn't been able to react with our Gold NPs. This step permits us to purify our solution so that we'll then be able to quantify labelled NPs using fluorescence spectroscopy. The first of our experiment is to lower the pH of our solution so that the reaction stops; to do so, we have to neutralize Tris molecules by using HCl (we have to add an amount of HCl which is equal to the amount of Tris we added yesterday). Protocol
DataCalculations for the volume of HCl needed to neutralize Tris. Concentration of Tris buffer = 50mM Concentration of HCl solution = 0.948M
Volume of Tris buffer = 321µL => Number of mole of Tris = 321 x 10^-6 x 50 x 10^-3 = 16µM => Volume of HCl solution to add = (1.6 x 10^-5)/0.948 = 16.8µL
Volume of Tris buffer = 1.5mL => Number of mole of Tris = 1.5 x 10^-3 x 50 x 10^-3 = 75µM => Volume of HCl solution to add = (7.5 x 10^-5)/0.948 = 79µL
Volume of Tris buffer = 600µL => Number of mole of Tris = 600 x 10^-6 x 50 x 10^-3 = 32µM => Volume of HCl solution to add = (3.2 x 10^-5)/0.948 = 32µL Notes
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