User:Matthew R Skorski/Notebook/471 - Exp BioChem/2015/11/10

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To perform Ocean Optics on Proteinase K at 10 nM concentration,


For instructions on the Ocean Optics protocol see 9/9/15.

  1. Preparing AuNP fiber samples
    1. Five 1 mL predried fiber samples were obtained
    2. 0.2mL of 100mM Tris and 50mM CaCl2 pH 8 buffer was added into the first eppendorf tube of fiber samples
    3. This was continued for all five samples
    4. All five samples were combined into the cuvette to run with the ocean optics
  2. Preparing Protinase K solution
    1. Proteinase K tube 1G was mixed with 1 mL of 100mM Tris and 50mM CaCl2 pH 8 buffer
    2. Proteinase K conentration: (0.00154g)*(1mol/28,900g)*(1/0.001L)= 0.0000532 M Protinase K
      1. Proteinase K dilution: (53278 nM)*(0.1 mL Proteinase K) = (x nM)*(10 mL total dilution) => x = 532 nM
      2. Proteinase K dilution: 0.1 mL of proteinase k solution was mixed with 9.9 mL of buffer
    3. Amount of Proteinase K solution needed for 3mL with 1μM concentration: M1*V1 = M2*V2 => (532M)*(V1) = (10 nM)*(3 mL) => V1 = 56.3 μL Protinase K
    4. Amount of Buffer solution need to get to 3mL: (3mL total)-(0.056 mL Protinase K solution) - 1mL fibers = 1.940 mL buffer
  3. The Protinase K solution, AuNP fiber samples, and buffer were mixed in a cuvette and run through the Ocean Optics