User:Sydney Marshall/Notebook/Protease Project/2015/09/09

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Objective

  • Utilize Ocean Optics spectrometer for UV-Vis Measurements of Proteinase K degradation
  • Prepare Proteinase K samples for future experiments

Methods

The procedure for today's lab is taken from Dr. Harting's notebook entry.

  • A 3mL solution of Proteinase K was made.
    • Mass of eppendorf tube = 1.01522g, Mass of Proteinase K = 0.00104g MW = 28,900g/mol
    • 0.00104g of Proteinase K was measured and placed in an eppendorf tube.
    • 1mL of water was added into the solution. The molarity of this solution was calculated using the following calculations: Proteinase Calc.JPG
    • A 3mL solution must be created. In order to do so, the following calculations were done:
      • M1V1=M2V2
      • M1=0.00003599M,M2=0.000001M,V2=0.003L, V1=?
      • (0.00003599M)*(V1)=(0.000001M)*(0.003L)
      • V1= 0.00008336L = 0.08336mL of current Proteinase K solution
    • The solution of Proteinase K will be composed of 0.08336mL of Proteinase Ksolution, 0.3mL of buffer, 1mL of fibers, thereby needing 1.617mL of distilled water to fulfill the total 3mL that is needed.
    • The 3mL solution was placed in a cuvette and measurements were taken using Ocean Optics. The 0.08336mL of proteinase K was added after the first 2 reading points.
  • 1mg stocks of Proteinase K were measured for future experiments.
    • The eppendorf tubes were also measured.
    • The tubes were labeled and stored in -20C for future uses. The table below shows the masses of each of the tubes that were stored in the freezer.

Data

Proteinase K Tube No. Eppendorf Tube Mass (g) Proteinase K Mass (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

Chem671 09032015 AMS.png
In the middle of the OceanOptics analysis, gold was nowhere to be found within the cuvette. A spectra is to be done soon.