User:Beatriz Gimenez De C./Notebook/572/2015/01/27

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Tasks

  • Produce the lysozyme AuNP fibers (Made 50, 5 mL samples)
  • Weigh 4 mg of Pepsine protease into 8 tubes
  • Prepare buffer

Procedure

  • Lysozyme-AuNP fibers preparation
    • Prepare Lysozyme 20 μM stock solution
      • Measured 0.0261 g of lysozyme into a 100 mL volumetric flask and filled the rest with distilled water
    • Prepare 2.5×10-3 M HAuCl4 stock solution
      • added 0.08539 g of HAuCl4 into a 100 mL volumetric flask and filled the rest with distilled water
    • Transfer 0.5 mL 2.5×10-3 M HAuCl4 into 50 culture tubes
      • 0.5×10-3(L)*2.5×10-3(mol/L)= 1.25 (mol) needed for a 45:1 ratio
    • Transfer 1.39 mL of lysozyme stock solution into 50 culture tubes
      • 2.77777778×10-8(mol)/20*10^-6(mol/ml)= 1.39 mL for a 45:1 ratio
    • Fill up 50 culture tubes to 5 mL total with distilled water
    • Placed in oven at 80ºC for 4h
  • Weighed Protease pepsin into 8 tubes, for the next experiment.
    • 1. 0.00530 g
    • 2. 0.00398 g
    • 3. 0.00391 g
    • 4. 0.00470 g
    • 5. 0.00419 g
    • 6. 0.00465 g
    • 7. 0.00780 g
    • 8. 0.00584 g
  • Prepared 5 mM Tris/0.2 M NaCl buffer for reaction of the synthesized lysozyme-AuNP fibers with the proteases
    • 1000 mL of buffer was prepared, used 11.68960 g of NaCl + 0.78922 g of Tris in approx. 1000 mL of distilled water.
    • 0.5 N HCl added until pH=8.067.