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:
- 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
|
In the middle of the OceanOptics analysis, gold was nowhere to be found within the cuvette. A spectra is to be done soon.
|