User:Paul Rothenberg/Notebook/CHEM 571 Fall 2014/2015/03/31: Difference between revisions
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== | ==New Reaction/Sampling Protocol== | ||
Here's the new protocol we developed for analysis | 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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==Notes from Protocol Prep== | ==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. | When trying to resuspend the fibers in solution, they were vortexed gently. The solution became colloidal (purple) in color with fibers suspended. When the supernatant was extracted out and replaced with buffer, the fibers did not reconstitute into solution as expected. The solution was sonicated; it appeared that the sonication possibly created a colloidal solution from the fibers. | ||
Instead, the fibers will be dispersed in the individual test tubes, and concentrated buffer added. This will prevent excess transferring of fibers and dispersion of fibers into colloid. | |||
'''The new protocol should be something similar to this:''' | |||
#Suspend fibers in individual glass tubes | |||
#Extract some amount of homogenous fiber solution and transfer to an epi-tube | |||
#Add concentrated buffer to bring the final buffer concentration to 50 mM Tris/30 mM CaCl<sub>2</sub> | |||
#Add protease solution to the mixture and begin shaking | |||
#When the reaction is finished, spin the individual tube for 30 seconds and extract the supernatant for analysis | |||
This should minimize the errors from the protocol we ran today. | |||
Revision as of 20:10, 31 March 2015
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New Reaction/Sampling ProtocolHere's the new protocol we developed for analysis:
Notes from Protocol PrepWhen trying to resuspend the fibers in solution, they were vortexed gently. The solution became colloidal (purple) in color with fibers suspended. When the supernatant was extracted out and replaced with buffer, the fibers did not reconstitute into solution as expected. The solution was sonicated; it appeared that the sonication possibly created a colloidal solution from the fibers. Instead, the fibers will be dispersed in the individual test tubes, and concentrated buffer added. This will prevent excess transferring of fibers and dispersion of fibers into colloid. The new protocol should be something similar to this:
This should minimize the errors from the protocol we ran today.
|