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

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Objective

To react proteinase K with AUNP protein fibers while measuring the absorbance with the Ocean Optics spectrometer.

Description

The overall protocol for the day can be found at Dr. Harting's notebook.

  1. 3mL Proteinase K solution with 1μM Proteinase K, 10mM Tris, 5mM CaCl2, and 1mL of AUNP protein fibers
    1. Measured out 0.00104g of proteinase K into an eppendorf tube (mass 1.01522g)
    2. Added 1mL of distilled water to the protein and shook to dissolve the protein
      1. Resulting molarity of Proteinase K solution: (0.00104g)*(1mol/28,900g)*(1/0.001L)=0.00003599M
      2. Amount of Proteinase K solution needed for 3mL with 1μM concentration: M1*V1=M2*V2 => (0.00003599)*(V1)=(0.000001)*(0.003) => V1=0.00008336L = 0.08336mL
      3. Amount of water required for 3mL: 3mL final - 0.3mL buffer - 0.08336mL Proteinase K - 1mL of AUNP fibers = 1.617 mL water
    3. Added 0.3mL of 100mM Tris and 50mM CaCl2 pH 7.4 buffer solution, 1mL of AUNP protein fibers, and 1.617mL of water to a cuvette
    4. Inserted the cuvette into the Ocean Optics for 2.5 hours with measurements every 2 minutes (other specs found here under Description). 0.083337mL of the Proteinase K solution was added after first two measurements were made.
  1. Measured out 1mg Proteinase K stock eppendorf tubes for future use
    1. The tubes were massed and tared before adding the proteinase
    2. Samples were numbered and stored in the freezer for future use

Results

Tube Number Mass of Tube (g) Mass of Proteinase-K (g)
1 1.02157 0.00085
2 1.03162 0.00087
3 1.02798 0.00095
4 1.03275 0.00114
5 1.03057 0.00123
6 1.01392 0.00067
7 1.02938 0.00118
8 1.02917 0.00123
9 1.02993 0.00085
10 1.02223 0.00127
11 1.02055 0.00107
12 1.03129 0.00103
13 1.01359 0.00121
14 1.02997 0.00129
15 1.03117 0.00116


2015 9 9 Reaction of Proteinase-K and Gold Nanoparticles.png WHAT DOES THIS GRAPH EVEN SHOW??????

Matt Hartings Can you make any observations about your ocean optics data? what happened?