User:Michael F. Nagle/Notebook/Chem 571/2012/09/19

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(Procedure)
(Procedure)
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*Tris added to Au/BSA
*Tris added to Au/BSA
**In order to determine how various concentrations of tris affect AuNPs in solution, tris was added to 70 [HAuCl<sub>4</sub>/BSA] solutions prepared [[User:Michael_F._Nagle/Notebook/Chem_571/2012/09/12|last week]] at .05mM, .5mM, 5mM, 50mM, 100mM, 200mM, 500mM and 1M. These [[User:Michael_F._Nagle/Notebook/Chem_571/2012/09/25|are to be analyzed by UV/Vis]].
**In order to determine how various concentrations of tris affect AuNPs in solution, tris was added to 70 [HAuCl<sub>4</sub>/BSA] solutions prepared [[User:Michael_F._Nagle/Notebook/Chem_571/2012/09/12|last week]] at .05mM, .5mM, 5mM, 50mM, 100mM, 200mM, 500mM and 1M. These [[User:Michael_F._Nagle/Notebook/Chem_571/2012/09/25|are to be analyzed by UV/Vis]].
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**The following calculations were completed to determine how much tris was needed in each solution. [[User:Puja_Mody/Notebook/Chem_571:_Gold_Nanoparticles/2012/09/19|Puja Mody]] prepared the tris stock.
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***Stock solution for Tris was made
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**m<sub>1</sub>v<sub>1</sub>=m<sub>2</sub>v<sub>2</sub>
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****1mol/L *.025L = .025mol Tris needed
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**1000mM*6mL=1000mM*x
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****.025mol * 121.14g/mol = 3.0285g Tris weighed
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***6mL Tris stock for 1M solution
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***The following calculations were completed to determine how much tris was needed in each solution for concentrations 1M, 500mM, 200mM and 100mM. A serial dilution was done to prepare concentrations .05mM-50mM from the 500mM solution.
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**500mM*6mL=1000mM*x8
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***m<sub>1</sub>v<sub>1</sub>=m<sub>2</sub>v<sub>2</sub>
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***3mL needed for 500mM solution
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***1000mM*6mL=1000mM*x
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**200mM*6mL=1000mM*x
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****6mL Tris stock for 1M solution
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***1.2mL Tris stock for 200mM solution
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***200mM*6mL=1000mM*x
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**100mM*6mL=1000mM*x
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****1.2mL Tris stock for 200mM solution
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***.6mL Tris stock for 100mM solution
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***100mM*6mL=1000mM*x
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**50mM*6mL=1000mM*x
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****.6mL Tris stock for 100mM solution
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***.3mL Tris stock for 50mM solution
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**5mM*6mL=1000mM*x
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***.03mL Tris stock for 5mM solution
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**.5mM*6mL=1000mM*x
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***.003mL Tris stock for .5mM solution
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**.05mM*6mL=1000mM*x
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***.0003mL Tris stock for .05mM solution
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__NOTOC__
__NOTOC__

Revision as of 00:41, 7 December 2012

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Objectives

  • Begin growing E. Coli for ADA expression. The ADA made will be used to nucleate AuNPs
  • make elution and binding buffers to be used for extraction and purification of ADA
  • prepare solutions of tris in Au/BSA, to be analyzed by UV/Vis to determine how various concentrations affect AuNPs in solution

Procedure

  • Expressing ADA
    • Cultures were prepared and E. Coli with a plasmid for Adenosine Deaminase (ADA) were added and incubated overnight, by Mary Mendoza. She also prepared binding and elution buffers for extraction.
    • Binding and elution buffers were prepared by Puja Mody
  • Tris added to Au/BSA
    • In order to determine how various concentrations of tris affect AuNPs in solution, tris was added to 70 [HAuCl4/BSA] solutions prepared last week at .05mM, .5mM, 5mM, 50mM, 100mM, 200mM, 500mM and 1M. These are to be analyzed by UV/Vis.
      • Stock solution for Tris was made
        • 1mol/L *.025L = .025mol Tris needed
        • .025mol * 121.14g/mol = 3.0285g Tris weighed
      • The following calculations were completed to determine how much tris was needed in each solution for concentrations 1M, 500mM, 200mM and 100mM. A serial dilution was done to prepare concentrations .05mM-50mM from the 500mM solution.
      • m1v1=m2v2
      • 1000mM*6mL=1000mM*x
        • 6mL Tris stock for 1M solution
      • 200mM*6mL=1000mM*x
        • 1.2mL Tris stock for 200mM solution
      • 100mM*6mL=1000mM*x
        • .6mL Tris stock for 100mM solution
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