User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/23: Difference between revisions
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==ADA preparation== | ==ADA preparation== | ||
* All ADA procedures for this lab period was conducted by [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/23|Puja Mody]]. | * All ADA procedures for this lab period was conducted by [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/23|Puja Mody]]. The same approach was conducted during [[User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/09/19|09/19/2012]] | ||
* This was a two day procedure continued by Michael Nagle on the following day, [[User:Michael F. Nagle/Notebook/Chem 571/2012/10/23|10/23/2012]] | |||
* The main concept for the procedure: ''E. coli'' was transformed to express ADA. After a 4 h. protein expression on an incubator shaker, the solution were centrifuge. The supernatant was collected and stored since this contained the soluble ADA while the pellets (cellular debris) were discarded. | |||
==Au/lysozyme solutions== | ==Au/lysozyme solutions== | ||
* The calculations were supposed to be posted on the notebook of [[User:Michael F. Nagle/Notebook/Chem 571/2012/10/23|Michael Nagle]]. | * The calculations were supposed to be posted on the notebook of [[User:Michael F. Nagle/Notebook/Chem 571/2012/10/23|Michael Nagle]]. | ||
* The calculation the author had executed for stock solution preparation is on [[User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/17|10/17/2012]]. The lysozyme stock solution was 15.24 μM and the gold was at 8.24 mM. | |||
* It was decided that the lysozyme would be kept at a 1.5 μM concentration for all sample varying the Au concentration. The same mole ratios from Au/BSA were used 60, 80, 100, 120, 128, 130, 132, 133, 134, 136, 138, 140, 160, and 170. | |||
* When the Au/lysozyme solutions were made, Nagle placed the rack in the Thermo Scientific incubator at 50°C for 4h. | |||
==AAS of Au/BSA solution of == | ==AAS of Au/BSA solution of 10/17/12== | ||
* The Au/BSA solutions were taken out from the oven. Mole ratios 60, 80, and 120 were the only solutions that exhibited a consistent, clear purple color with no fibers. This was indicative that all gold nanoparticles were in solution and none have nucleated to BSA. | |||
* Mole ratios 100 and others produced fibers, having solutions clear and colorless. The liquid was taken from each solution and transferred to falcon tubes; leaving some liquid suspending the fibers in the original tube. As a result, they were irregular amounts on each tube. | |||
* The irregular amounts created difficulty for centrifugation. Hours was spent in balancing the liquid for each pair of tubes. | |||
* Time went by and the fibers formed pellets on the bottom of the tubes. The formation of pellets was acceptable to allow to take AAS of the solutions. | |||
* The program for the Shimadzu AA-6200 Atomic Absorption Flame Emission was set on autozero. The flame was ignited exhibiting a blue-green color. | |||
* The blank used for the experiment was water. | |||
* The standards were run followed by an interval of sample-blank measurements. The calibration curve can be seen on the data section of [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/23|Puja Mody's notebook.]] | |||
Revision as of 07:55, 7 December 2012
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ADA preparation
Au/lysozyme solutions
AAS of Au/BSA solution of 10/17/12
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