User:Beatriz Gimenez De C./Notebook/572/2015/02/11: Difference between revisions

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** 250 μL samples of each proteinase K reaction at 30 min time points + with 200 μL of 1:4 Bradford:Tris/NaCl + 1550 μL of Tris/NaCl buffer.
** 250 μL samples of each proteinase K reaction at 30 min time points + with 200 μL of 1:4 Bradford:Tris/NaCl + 1550 μL of Tris/NaCl buffer.
*Ocean Optics will be run at 37°C, scanning every 5 minutes for 6 hours.
*Ocean Optics will be run at 37°C, scanning every 5 minutes for 6 hours.
== Results==
*Ocean Optics
[[Image:1µM_ProtK_Kinetics_Chart.png|500px]]
[[Image:1µM_ProtK_Kinetics_AbsvsTime_Chart.png|500px]]
* Bradford
[[Image:Proteinase_K_Bradford_1uM.png]]
[[Image:Proteinase_K_Bradford_100_nM.png]]
[[Image:Proteinase_K_Bradford_10nM.png]]
[[Image:Proteinase_K_Bradford_1nM.png]]


==Note==
==Note==

Revision as of 21:48, 5 May 2015

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Tasks

  • Create a Bradford calibration curve with Lysozyme.
    • 250 μL samples of each proteinase K reaction at 30 min time points + with 200 μL of 1:4 Bradford:Tris/NaCl + 1550 μL of Tris/NaCl buffer.
  • Ocean Optics will be run at 37°C, scanning every 5 minutes for 6 hours.

Results

  • Ocean Optics

  • Bradford

Note

  • Used incorrect stock solution for protease solution. The actual concentration is 1 nM while the expected concentration was 1 µM of protease.
  • The experiment was repeated with the same preparation method for the fibers,
    • 69 µL of protease solution, made from 10 µL protease stock + 990 µL buffer, were diluted with additional 2.931 mL of buffer. [ for 10nM protease concentration]
    • 6.9 µL of protease solution, made from 1 µL protease stock + 999 µL buffer, were diluted with 2.993 mL of buffer. [for 1 nM protease concentration]
  • Ocean Optics were run at 37°C, scanning every 5 minutes for 6 hours.



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