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

From OpenWetWare
Jump to: navigation, search
128px Project name Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png


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


  • 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)


  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

16nov-GoldNPs result.jpg