- 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
- Prepare AuNP fibers - vortex each 5 mL test tube containing fibers and combine some necessary amount into a large falcon tube.
- Centrifuge the falcon tube allowing the fibers to settle at the bottom and pour off the supernatant liquid.
- Add Tris/CaCl2 buffer and vortex again, resulting in a homogenous solution of fibers which can be equipartitioned between 20 sampling epi-tubes.
- Add protease to each epi tube. (1 epi-tube per each timepoint being sampled)
- 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.
- Collect the supernatant for analysis.
- 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.