User:Floriane Briere/Notebook/CHEM-496/2011/11/16

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Objective

Today we are going to continue protein expression and keep investigating on gold NPs.

Especially, we are going to investigate how the BSA/Gold ratio affects the reaction. We are going to use 4 solutions:

  1. Tube 1: ratio of 0.35/100 (BSA/Gold); identical to the ratio we used when we obtained purple fibers; with a 0.1ml BSA (15.4µM) + 0.1ml HAuCl4 (4.3mM) into a 1ml solution.
  2. Tube 2: ratio of 0.5/100; identical to the ratio we used when we obtained a purple solution; with 0.97ml BSA (15.5µM) + 0.58ml HAuCl4 (4.3mM) into a 10ml solution
  3. Tube 3: ration of 0.5/80 to test the effect of Gold decrease
  4. Tube 4B: ration of 0.5/60 to test the effect of Gold decrease

Protocol

  • Gold NPs synthesis:
  1. Prepare 4 different tubes to test the consequences of a modification of the BSA/Gold ratio:
    1. Tube 1: 0.1ml BSA (30µM) + 0.2ml HAuCl4(4.17mM) + 0.7ml water
    2. Tube 2: 0.1ml BSA (30µM) + 0.143ml HAuCl4 (4.17mM) + 0.757ml water
    3. Tube 3: 0.1ml BSA (30µM) + 0.115ml HAuCl4 (4.17mM) + 0.785ml water
    4. Tube 4A: 0.1ml BSA (30µM) + excess of HAuCl4
    5. Tube 4B: 0.1ml BSA (30µM) + 0.086ml HAuCl4 (4.17mM) + 0.814ml water
  2. Heat the tubes at 80°C in the heatblock for 2 hours (constant temperature)
  • Protein expresion:
  1. Add to each medium which has been prepared yesterday:
    1. 1ml of Ampicillin (300mg Ampicilin + 4ml water)
    2. 1ml of cell suspension (which grew up overnight)
    3. 1ml of IPTG (Isopropyl β-D-1-thiogalactopyranoside) (380mg + 4ml water)
  2. Place the 4 preparations in the shaker for several hours to let the cells grow up
  1. Centrifuge the 4 preparations
  2. Add medium to the cells and place the preparation into the fridge (-80°C)

Results

  1. Tube 1: Clear solution and yellow fibers
  2. Tube 2: Clear solution and purple fibers
  3. Tube 3: purple solution
  4. Tube 4A: The solution remained yellow
  5. Tube 4B: Clear solution and yellow fibers



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