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

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## Cultures were prepared and E. Coli with a plasmid for Adenosine Deaminase (ADA) were added and incubated overnight, by [[User:Mary_Mendoza/Notebook/CHEM_571_Experimental_Biological_Chemistry_I/2012/09/19|Mary Mendoza]]. She also prepared binding and elution buffers for extraction.
## Cultures were prepared and E. Coli with a plasmid for Adenosine Deaminase (ADA) were added and incubated overnight, by [[User:Mary_Mendoza/Notebook/CHEM_571_Experimental_Biological_Chemistry_I/2012/09/19|Mary Mendoza]]. She also prepared binding and elution buffers for extraction.


 
#In order to determine how various concentrations of Tris affect the formation of AuNP by BSA, tris buffer solution was added to HAuCl<sub>4</sub> and BSA solutions prepared [[User:Michael_F._Nagle/Notebook/Chem_571/2012/09/12|last week]] at molarities of .05mM, .5mM, 5mM, 50mM, 100mM, 200mM, 500mM and 1M. They are to be analyzed by UV/Vis.
#In order to determine how various concentrations of Tris affect the formation of AuNP by BSA, tris buffer solution was added to HAuCl<sub>4</sub> and BSA tubes at molarities of .05mM, .5mM, 5mM, 50mM, 100mM, 200mM, 500mM and 1M. They are to be analyzed by UV/Vis.
##m<sub>1</sub>v<sub>1</sub>=m<sub>2</sub>v<sub>2</sub>
##m<sub>1</sub>v<sub>1</sub>=m<sub>2</sub>v<sub>2</sub>
##500mM*6mL=1000mM*x8##3mL
##500mM*6mL=1000mM*x8##3mL
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##.05mM*6mL=1000mM*x
##.05mM*6mL=1000mM*x
##.0003mL Tris stock for .05mM solution
##.0003mL Tris stock for .05mM solution
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 11:34, 7 October 2012 (EDT)''':why are you making these buffers? how much as added to what concentrations of HAuCl4 nad BSA? is this before or after AuNP formation?


==Observations==
==Observations==

Revision as of 20:16, 25 October 2012

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Objectives

  • begin protein expression
  • make elution and binding buffers
  • collect E. Coli with ADA

Procedure

  1. Expressing ADA
    1. 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.
  1. In order to determine how various concentrations of Tris affect the formation of AuNP by BSA, tris buffer solution was added to HAuCl4 and BSA solutions prepared last week at molarities of .05mM, .5mM, 5mM, 50mM, 100mM, 200mM, 500mM and 1M. They are to be analyzed by UV/Vis.
    1. m1v1=m2v2
    2. 500mM*6mL=1000mM*x8##3mL
    3. 1000mM*6mL=1000mM*x
    4. 6mL Tris stock for 1M solution
    5. 200mM*6mL=1000mM*x
    6. 1.2mL Tris stock for 200mM solution
    7. 100mM*6mL=1000mM*x
    8. .6mL Tris stock for 100mM solution
    9. 50mM*6mL=1000mM*x
    10. .3mL Tris stock for 50mM solution
    11. 5mM*6mL=1000mM*x
    12. .03mL Tris stock for 5mM solution
    13. .5mM*6mL=1000mM*x
    14. .003mL Tris stock for .5mM solution
    15. .05mM*6mL=1000mM*x
    16. .0003mL Tris stock for .05mM solution

Observations

One of the flasks with broth appeared a different shade of yellow this morning prior to centrifuging.

  • Abigail E. Miller 11:34, 7 October 2012 (EDT):why is this important? did this change your procedure?


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