User:Beatriz Gimenez De C./Notebook/572/2015/04/01

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Tasks

  • New protocol
    • Vortex each 5 mL test tube of fibers until solution seems homogeneous
    • Prepare concentrated buffer -> 50 mM Tris-HCl/20 mM CaCl2 pH 8 buffer.
    • Prepare Proteinase K solution
    • Add 0.927 mL of the homogeneous fiber sample from the vortexed 5 mL test tube + 0.020 mL of a 2.5 M Tris-HCl/0.5 M CaCl2 pH 8 buffer + 0.053 mL of protease for a total volume of 1 mL into an epitub.
    • When the reaction is finished, spin the individual tube for 30 seconds and extract the supernatant for analysis
  • Run Bradford of Proteinas K for 0, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 15, 20, 25, 30, 40, 50 ,60 min.
    • 200μM of a 1 in 4 Bradford dilution (1 mL Bradford in 3 mL of Tris/NaCl buffer)
    • 150μM of each proteinase K sample
    • 550μM of tris/NaCl buffer

Results

Bradford analysis new method.png