User:Matthew R Skorski/Notebook/471 - Exp BioChem/2015/09/30

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Objective

To perform Ocean Optics on Proteinase K at 1 μM concentrations as well as run the blanks for the Bradford reaction from yesterday.

Description

For instructions on the Ocean Optics protocol see 9/9/15. For Bradford see 9/29/15

  1. Preparing AuNP fiber samples
    1. Five 1 mL predried fiber samples were obtained
    2. 1mL of 100mM Tris and 50mM CaCl2 pH 7.4 buffer was added into the first eppendorf tube of fiber samples and then the entire contents of the tube were removed and added to the second sample
    3. This was continued for all five samples
  2. Preparing Protinase K solution
    1. Protinase K tube 8 was mixed with 1 mL of 100mM Tris and 50mM CaCl2 pH 7.4 buffer
    2. Protinase K conentration: (0.00123g)*(1mol/28,900g)*(1/0.001L)= 0.0000426 M Protinase K
    3. Amount of Proteinase K solution needed for 3mL with 1μM concentration: M1*V1 = M2*V2 => (42.6 μM)*(V1) = (1 μM)*(3 mL) => V1 = 70.5 μL Protinase K
    4. Amount of Buffer solution need to get to 3mL: (3mL total)-(0.0705 mL Protinase K solution) - 1mL fibers = 1.630 mL buffer
  3. The Protinase K solution, AuNP fiber sample, and buffer were mixed in a cuvette and run through the Ocean Optics
  4. Running blanks for Bradford
    1. Amount of Proteinase K solution needed for 1mL with 1μM concentration: M1*V1 = M2*V2 => (42.6 μM)*(V1) = (1 μM)*(1 mL) => V1 = 23.5 μL Protinase K
    2. Amount of Buffer solution need to get to 1mL: (1mL total)-(0.0235 mL Protinase K solution) = 0.9765 mL buffer
    3. Tubes were incubated for identical times to previous day's work

Results

OCEAN OPTICS DATA NEEDS TO BE WORKED UP