User:Sydney Marshall/Notebook/Protease Project/2015/09/30

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Objective

  • Perform OceanOptics on Proteinase-K at 1 μM concentration
  • Run Bradford blanks from yesterday's experiment

Description

Protocol for OceanOptics Experiment can be found here. Protocol for Bradford can be found here.

  • Preparing AuNP fiber samples
    • Five 1 mL predried fiber samples were obtained
    • 1mL of 100mM Tris/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

This was continued for all five samples

  • Preparing Proteinase K solution
    • Proteinase K (tube 8) was mixed with 1 mL of 100mM Tris/50mM CaCl2 pH 7.4 buffer
    • Protinase K conentration: (0.00123g)*(1mol/28,900g)*(1/0.001L)= 0.0000426 M Proteinase K
    • 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
    • Amount of Buffer solution need to get to 3mL: (3mL total)-(0.0705 mL Protinase K solution) - 1mL fibers = 1.630 mL buffer
  • Proteinase K solution was run in cuvette through OceanOptics
  • Running blanks for Bradford
    • 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
    • Amount of Buffer solution need to get to 1mL: (1mL total)-(0.0235 mL Protinase K solution) = 0.9765 mL buffer
    • Tubes were incubated for identical times to previous day's work


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