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

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Procedure)
(Procedure)
Line 21: Line 21:
****.025mol * 121.14g/mol = 3.0285g Tris weighed
****.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.
***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.
-
***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>
-
***1000mM*6mL=1000mM*x
+
****1000mM*6mL=1000mM*x
-
****6mL Tris stock for 1M solution
+
*****6mL Tris stock for 1M solution
-
***200mM*6mL=1000mM*x
+
****200mM*6mL=1000mM*x
-
****1.2mL Tris stock for 200mM solution
+
*****1.2mL Tris stock for 200mM solution
-
***100mM*6mL=1000mM*x
+
****100mM*6mL=1000mM*x
-
****.6mL Tris stock for 100mM solution
+
*****.6mL Tris stock for 100mM solution
__NOTOC__
__NOTOC__

Revision as of 00:41, 7 December 2012

Project name Main project page
Previous entry      Next entry

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
Personal tools