User:Paul Rothenberg/Notebook/CHEM 571 Fall 2014/2015/03/31: Difference between revisions

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==Entry title==
=='''New Reaction/Sampling Protocol'''==
* Insert content here...
 
#Prepare AuNP fibers - vortex each 5 mL test tube containing fibers and combine some necessary amount into a large falcon tube.
 
#Centrifuge the falcon tube allowing the fibers to settle at the bottom and pour off the supernatant liquid.
 
#Add Tris/CaCl<sub>2</sub> buffer and vortex again, resulting in a homogenous solution of fibers which can be equipartitioned between 20 sampling epi-tubes.
 
#Add protease to each epi tube. (1 epi-tube per each timepoint being sampled)
 
#At 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60 min stop the reaction by removing from the "incubator/shaker" and centrifuging for 30 sec.
 
#Collect the supernatant for analysis.
 
Here's the new protocol we developed for analysis.
 
==Notes from Protocol Prep==
 
#When trying to resuspend the fibers in solution, they were vortexed gently. The solution became colloidal in color with fibers suspended.  




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New Reaction/Sampling Protocol

  1. Prepare AuNP fibers - vortex each 5 mL test tube containing fibers and combine some necessary amount into a large falcon tube.
  1. Centrifuge the falcon tube allowing the fibers to settle at the bottom and pour off the supernatant liquid.
  1. Add Tris/CaCl2 buffer and vortex again, resulting in a homogenous solution of fibers which can be equipartitioned between 20 sampling epi-tubes.
  1. Add protease to each epi tube. (1 epi-tube per each timepoint being sampled)
  1. At 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60 min stop the reaction by removing from the "incubator/shaker" and centrifuging for 30 sec.
  1. Collect the supernatant for analysis.

Here's the new protocol we developed for analysis.

Notes from Protocol Prep

  1. When trying to resuspend the fibers in solution, they were vortexed gently. The solution became colloidal in color with fibers suspended.



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