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

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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