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

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