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

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(Procedure)
(Procedure)
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****100mM*6mL=1000mM*x
****100mM*6mL=1000mM*x
*****.6mL Tris stock for 100mM solution
*****.6mL Tris stock for 100mM solution
 +
**The contents of each tube were as follows:
 +
|+ Amount of Tris stock and water in each tube
 +
! scope="col" |'''Tris Buffer concentration'''
 +
! scope="col" |'''amount of water (mL)'''
 +
! scope="col" |'''amount of tris (mL)'''
 +
|-
 +
| .05 mM||0.9938||0.0625
 +
|-
 +
| .5 mM||0.9875||0.0125
 +
|-
 +
| 5 mM||0.975||0.025
 +
|-
 +
| 50 mM||0.95||0.05
 +
|-
 +
| 100 mM||0.9||0.1
 +
|-
 +
| 200 mM||0.8||0.2
 +
|-
 +
| 500 mM||0.5||0.5
 +
|-
 +
| 1 M||0||1
 +
|}
__NOTOC__
__NOTOC__

Revision as of 23:43, 6 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.
      • 1M 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
    • The contents of each tube were as follows:
Amount of Tris stock and water in each tube
Tris Buffer concentration amount of water (mL) amount of tris (mL)
.05 mM0.99380.0625
.5 mM0.98750.0125
5 mM0.9750.025
50 mM0.950.05
100 mM0.90.1
200 mM0.80.2
500 mM0.50.5
1 M01
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