User:Beatriz Gimenez De C./Notebook/572/2015/03/31

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Tasks

  • Preparation of more AuNP Fibers
    • Prepare Lysozyme 20 μM stock solution
      • Measured 0.01439 g of lysozyme into a 50 mL volumetric flask and filled the rest with distilled water
    • Prepare 2.5×10-3 M HAuCl4 stock solution
      • added 0.04361 g of HAuCl4 into a 50 mL volumetric flask and filled the rest with distilled water
    • Transfer 0.5 mL 2.5×10-3 M HAuCl4 into 29 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 29 culture tubes to 5 mL total with distilled water
    • Placed in oven at 80ºC for 4h
  • Prepare concentrated Tris/NaCl buffer (2.5 M Tris + 0.5 M NaCl) * Proteinase K kinetics with new protocol
  1. Prepare AuNP fibers - vortex each 5 mL test tube containing fibers and combine some necessary amount into a large falcon tube.
  2. Centrifuge the falcon tube allowing the fibers to settle at the bottom and pour off the supernatant liquid.
  3. Add Tris/CaCl2 buffer and vortex again, resulting in a homogenous solution of fibers which can be equipartitioned between 20 sampling epi-tubes.
  4. Add protease to each epi tube. (1 epi-tube per each timepoint being sampled)
  5. At 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60 min stop the reaction by removing from the "incubator/shaker" and centrifuging for 30 sec.
  6. Collect the supernatant for analysis.

Note

  • Protocol did not work, the AuNP fibers would not re-suspend in solution with the buffer.
  • New protocol was design for Wednesday.
    • Fibers will be dispersed in the individual test tubes + concentrated buffer
    • Preventing excess transfer of fibers and maintaining the fibers in solution.




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