User:Floriane Briere/Notebook/CHEM-496/2011/10/25: Difference between revisions
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## The stacking gel: compare to the resolving gel, the stacking gel has a lower concentration of acrymalide and a lower pH. It allows to concentrate the proteins into a thight band. | ## The stacking gel: compare to the resolving gel, the stacking gel has a lower concentration of acrymalide and a lower pH. It allows to concentrate the proteins into a thight band. | ||
## The resolving gel: it permits to separate the proteins which are going to migrate through the gel according to their molecular weight. | ## The resolving gel: it permits to separate the proteins which are going to migrate through the gel according to their molecular weight. | ||
The protocol was esthablished using this [http://ergo.berkeley.edu/be115/protean3.pdf manuel]. | |||
==Protocol== | ==Protocol== | ||
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## Load the sample into the wells | ## Load the sample into the wells | ||
## Run the gel for 35 minutes at 200 volts | ## Run the gel for 35 minutes at 200 volts | ||
# Stain the polyacrylamide gel: | |||
## Cover the gel with stain (blue) | |||
## Leave overnight on the shaker | |||
[[Category:Course]] | [[Category:Course]] |
Revision as of 09:33, 26 October 2011
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ObjectiveToday's experiment consists on performing a gel electrophoresis on MPB-intein fusion which have been expressed and purified by groups 2 and 3. This experiment will help us to determine whether or not the expressions and purifications worked well. To do so, we are going to run a SDS-PAGE using a 12% discontinuous polyacrylamide gel.
The protocol was esthablished using this manuel. Protocol
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