User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/04: Difference between revisions

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==Procedure==
==Procedure==
* The absorbance of each sample were taken by UV-vis spectrophotometer. Graph were made with absorbance versus wavelength. See [[User:Dhea Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/09/04|Dhea Patel's Lab Notebook on 2012/09/04]] for details.
* The absorbance of each sample were taken by UV-vis spectrophotometer. Data with absorbance versus wavelength were obtained. The original sets of Au/BSA solution failed to work, and thus new sets of solution made by Dr. Abigail Miller were used for UV-vis spectrum. See [[User:Dhea Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/09/04|Dhea Patel's Lab Notebook on 2012/09/04]] for details regarding the appearance of unsuccessful solutions made last week and reacted solutions made by Dr. Abigail Miller.  
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 10:45, 17 September 2012 (EDT)''':shoe the data in your notebook and clearly explain what can be found in Dhea's notebook.
* Solutions of Au/BSA at various ratios were remade at newly calculated amounts. See [[User:Dhea Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/09/04|Dhea Patel's Lab Notebook on 2012/09/04]] for the details on each amount and concentration of HAuCl4 and BSA stock solution was used for each Au/BSA ratio. New stock solutions were remade. Solutions were wrapped around with aluminum foil and placed in incubator at 85°C for five hours. Incubator were cooled to room temperature until the next day.
* Solutions of Au/BSA at various ratios were remade. New stock solutions were remade. Solutions were wrapped around with aluminum foil and placed in incubator at 85°C for five hours. Incubator were cooled to room temperature until the next day.
* Theoretical amount of 0.1M Tris buffer with original formula weight of 121.14g/mol were roughly calculated by weighted out with 12.114g of tris mixed with 100mL of water. For details of the calculation, please refer to [[User:Melissa Novy/Notebook/CHEM-571/2012/09/04|Melissa's Notebook]] for the calculation of 0.1M tris buffer solution. Two sets of solutions were made. In each 100mL tris buffer, each sample was titrated to pH 8.0 and pH 10.0 with the addition of 1M HCl, respectively.
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 10:45, 17 September 2012 (EDT)''':how? what masses? what volumes where used? what ratios were prepared? I cannot recreate your experiments from what you have listed.
* New sets of Au/BSA solutions at various ratios that were pre-made on 2012/09/04 before were placed in centrifuge and were attempted to spin at 4000rpm for 5 minutes in room temperature. However, two glass test tubes that were placed on the outskirts of centrifuge shattered during the process. Glass tubes broke during the process of centrifugation and were put to halt as a result. All solutions were then transferred into falcon tubes. The Au/BSA solution at ratio 130 and 128 were disposed due to mix of broken glass. Some purple fibers were stuck on glass test tubes while being transferred to falcon tube and are lost during the transfer.
* 1.2114g of Tris buffer was mixed with 100mL of distilled water to make 100mM of tris buffer solution. Two sets of solutions were made. In each 100mL tris buffer, each sample was titrated to pH 8.0 and pH 10.0 with the addition of 1M HCl, respectively.
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 10:45, 17 September 2012 (EDT)''':what were the actual masses for tris for each solution? show the calculation. what is the theoretical amount for a 100mM tris solution?
* New sets of Au/BSA solutions at various ratios that were pre-made before were placed in centrifuge and were attempted to spin at 4000rpm for 5 minutes in room temperature. However, two glass test tubes that were placed on the outskirts of centrifuge shattered during the process. All solutions were then transferred into falcon tubes. The Au/BSA solution at ratio 130 and 128 were disposed due to mix of broken glass. Some purple fibers were stuck on glass test tubes while being transferred to falcon tube and are lost during the transfer.
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 10:45, 17 September 2012 (EDT)''': did you recentrifuge or store the solutions in the falcon tubes? what solutions are these? from what day? what ratios? be more specific.


==Notes==
==Notes==
* The original sets of solutions did not form nano-particles, solutions appeared yellow in color. It was predicted that the HAuCl4 reacted with gold specula during the transfer.  
* The original sets of solutions did not form nano-particles, solutions appeared yellow in color. It was predicted that the HAuCl4 reacted with gold specula during the transfer.  
* A new set of solutions with different Au/BSA ratios were made beforehand at the same ratio and concentrations. For details of the concentrations, see [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/08/29|Notebook from 2012/08/29]]
* A new set of solutions with different Au/BSA ratios were made beforehand at the exact same ratio and concentrations. It was hypothesized that the spectula used during BSA weighting interacted with the BSA and thus undergo structural confirmation. This allowed the BSA stock to be inefficient to interact with the HAuCl4. As a result, the concentration of solutions for Au/BSA ratios were not changed. For details of the concentrations, see [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/08/29|Notebook from 2012/08/29]]
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 10:48, 17 September 2012 (EDT)''':did you use exactly the same stock solutions so all the volumes are precisely the same? what did you do with these solutions? what did you learn from them?
* The new set of solutions with different ratios of Au and BSA made in 2012/08/29 appeared purple in color. The 60 Au/BSA solution appeared cloudy and white. The 80 Au/BSA solution appeared as a purpose homogenous solution. The Au/BSA solution with other ratios are transparent with purple fiber formations.  
* The new set of solutions with different ratios of Au and BSA appeared purpose in color. The 60 Au/BSA solution appeared cloudy and white. The 80 Au/BSA solution appeared as a purpose homogenous solution. The Au/BSA solution with other ratios are transparent with purpose fiber formations.  
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 10:48, 17 September 2012 (EDT)''':what different ratios? '''purple''' in color? where did this set of solutions come from? what day? what concentrations? how were they prepared?




Revision as of 08:53, 5 October 2012

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Purpose

  • To run UV-Vis analysis on Au/BSA solutions at various ratios.
  • To remake solutions that did not give expected results.
  • To make Tris Buffer at 100mM at pH 8.0 and 10.0

Procedure

  • The absorbance of each sample were taken by UV-vis spectrophotometer. Data with absorbance versus wavelength were obtained. The original sets of Au/BSA solution failed to work, and thus new sets of solution made by Dr. Abigail Miller were used for UV-vis spectrum. See Dhea Patel's Lab Notebook on 2012/09/04 for details regarding the appearance of unsuccessful solutions made last week and reacted solutions made by Dr. Abigail Miller.
  • Solutions of Au/BSA at various ratios were remade at newly calculated amounts. See Dhea Patel's Lab Notebook on 2012/09/04 for the details on each amount and concentration of HAuCl4 and BSA stock solution was used for each Au/BSA ratio. New stock solutions were remade. Solutions were wrapped around with aluminum foil and placed in incubator at 85°C for five hours. Incubator were cooled to room temperature until the next day.
  • Theoretical amount of 0.1M Tris buffer with original formula weight of 121.14g/mol were roughly calculated by weighted out with 12.114g of tris mixed with 100mL of water. For details of the calculation, please refer to Melissa's Notebook for the calculation of 0.1M tris buffer solution. Two sets of solutions were made. In each 100mL tris buffer, each sample was titrated to pH 8.0 and pH 10.0 with the addition of 1M HCl, respectively.
  • New sets of Au/BSA solutions at various ratios that were pre-made on 2012/09/04 before were placed in centrifuge and were attempted to spin at 4000rpm for 5 minutes in room temperature. However, two glass test tubes that were placed on the outskirts of centrifuge shattered during the process. Glass tubes broke during the process of centrifugation and were put to halt as a result. All solutions were then transferred into falcon tubes. The Au/BSA solution at ratio 130 and 128 were disposed due to mix of broken glass. Some purple fibers were stuck on glass test tubes while being transferred to falcon tube and are lost during the transfer.

Notes

  • The original sets of solutions did not form nano-particles, solutions appeared yellow in color. It was predicted that the HAuCl4 reacted with gold specula during the transfer.
  • A new set of solutions with different Au/BSA ratios were made beforehand at the exact same ratio and concentrations. It was hypothesized that the spectula used during BSA weighting interacted with the BSA and thus undergo structural confirmation. This allowed the BSA stock to be inefficient to interact with the HAuCl4. As a result, the concentration of solutions for Au/BSA ratios were not changed. For details of the concentrations, see Notebook from 2012/08/29
  • The new set of solutions with different ratios of Au and BSA made in 2012/08/29 appeared purple in color. The 60 Au/BSA solution appeared cloudy and white. The 80 Au/BSA solution appeared as a purpose homogenous solution. The Au/BSA solution with other ratios are transparent with purple fiber formations.
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