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Team:Bordeaux/Results/Purification
From 2014.igem.org
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Cells are lysed and the protein mixture is collected. The ELP is purified by ITC (inverse transition cycling) | Cells are lysed and the protein mixture is collected. The ELP is purified by ITC (inverse transition cycling) | ||
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Method overview: | Method overview: | ||
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{{:Team:Bordeaux/Pied}} | {{:Team:Bordeaux/Pied}} | ||
Revision as of 02:23, 18 October 2014
Cells are lysed and the protein mixture is collected. The ELP is purified by ITC (inverse transition cycling)
Method overview:
- 2ml of polyethylenimine is added to the lysate, the mixture is centrifuged at 14,000rpm for 15min at 4°C.
- The supernatant is saved and brought to room temperature. NaCl (1 to 3M) is added while homogenising the solution.
- The mixture is then centrifuged at 14,000 rpm for 15min at 37°C.
- Supernatent is discarded and pellet is resuspended with 2ml of cold PBS.
- First and second steps are repeated but NaCl 5M is added so the ELP precipitate.
- Third and fourth steps are repeated.
- A final centrifugation is performed at 4°C for 10min (14,000rpm)
- The solution is dialysed and the samples are lyophilized.
