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Team:Cambridge/Experiments/Protein Purification
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Bacteria expressing his-tagged reflectin were lysed, and the protein was purified using a his-trap column and a denaturing protocol in order to solubilise reflectin. | Bacteria expressing his-tagged reflectin were lysed, and the protein was purified using a his-trap column and a denaturing protocol in order to solubilise reflectin. | ||
| - | + | ==Practice== | |
| + | *After high copy expression plasmids for his-tagged reflectin were successfully [assembled] and [transformed] in E. coli, cultures were incubated overnight with () arabinose to induce reflectin expression. | ||
| + | *[Buffers] were prepared, and their pH checked and readjusted if necessary on the day of purification. | ||
| + | *An [inclusion body prep] was performed with 50ml of overnight culture. | ||
Revision as of 10:50, 21 August 2011
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Protein Purification
Bacteria expressing his-tagged reflectin were lysed, and the protein was purified using a his-trap column and a denaturing protocol in order to solubilise reflectin.
Practice
- After high copy expression plasmids for his-tagged reflectin were successfully [assembled] and [transformed] in E. coli, cultures were incubated overnight with () arabinose to induce reflectin expression.
- [Buffers] were prepared, and their pH checked and readjusted if necessary on the day of purification.
- An [inclusion body prep] was performed with 50ml of overnight culture.
