User:Beatriz Gimenez De C./Notebook/572/2015/03/31: Difference between revisions
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** Fill up 29 culture tubes to 5 mL total with distilled water | ** Fill up 29 culture tubes to 5 mL total with distilled water | ||
** Placed in oven at 80ºC for 4h | ** Placed in oven at 80ºC for 4h | ||
* Prepare concentrated Tris/NaCl buffer (2.5 M Tris + 0.5 M NaCl) | |||
* Prepare concentrated Tris/NaCl buffer (2.5 M Tris + 0.5 M NaCl) * Proteinase K kinetics with new protocol | |||
#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. | |||
==Note== | |||
* Protocol did not work, the AuNP fibers would not re-suspend in solution with the buffer. | |||
* New protocol was design for Wednesday. | |||
** Fibers will be dispersed in the individual test tubes + concentrated buffer | |||
** Preventing excess transfer of fibers and maintaining the fibers in solution. | |||
Revision as of 13:20, 1 April 2015
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