To react proteinase K with AUNP protein fibers while measuring the absorbance with the Ocean Optics spectrometer.
The overall protocol for the day can be found at Dr. Harting's notebook.
- 3mL Proteinase K solution with 1μM Proteinase K, 10mM Tris, 5mM CaCl2, and 1mL of AUNP protein fibers
- Measured out 0.00104g of proteinase K into an eppendorf tube (mass 1.01522g)
- Added 1mL of distilled water to the protein and shook to dissolve the protein
- Resulting molarity of Proteinase K solution: (0.00104g)*(1mol/28,900g)*(1/0.001L)=0.00003599M
- 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
- Amount of water required for 3mL: 3mL final - 0.3mL buffer - 0.08336mL Proteinase K - 1mL of AUNP fibers = 1.617 mL water
- 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
- 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.
- Measured out 1mg Proteinase K stock eppendorf tubes for future use
- The tubes were massed and tared before adding the proteinase
- Samples were numbered and stored in the freezer for future use
| Tube Number
||Mass of Tube (g)
||Mass of Proteinase-K (g)
WHAT DOES THIS GRAPH EVEN SHOW??????
Matt Hartings Can you make any observations about your ocean optics data? what happened?