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

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=='''New Reaction/Sampling Protocol'''==
=='''New Reaction/Sampling Protocol'''==
Here's the new protocol we developed for analysis.


#Prepare AuNP fibers - vortex each 5 mL test tube containing fibers and combine some necessary amount into a large falcon tube.
#Prepare AuNP fibers - vortex each 5 mL test tube containing fibers and combine some necessary amount into a large falcon tube.
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#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.  
#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.
#Collect the supernatant for analysis.
Here's the new protocol we developed for analysis.


==Notes from Protocol Prep==
==Notes from Protocol Prep==

Revision as of 09:34, 31 March 2015

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New Reaction/Sampling Protocol

Here's the new protocol we developed for analysis.

  1. Prepare AuNP fibers - vortex each 5 mL test tube containing fibers and combine some necessary amount into a large falcon tube.
  2. Centrifuge the falcon tube allowing the fibers to settle at the bottom and pour off the supernatant liquid.
  3. Add Tris/CaCl2 buffer and vortex again, resulting in a homogenous solution of fibers which can be equipartitioned between 20 sampling epi-tubes.
  4. Add protease to each epi tube. (1 epi-tube per each timepoint being sampled)
  5. 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.
  6. Collect the supernatant 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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