Team:Bordeaux/Results/Purification

From 2014.igem.org

• 
• 
• 
• 
• 
• 
• 
• 
• 
• 
• 
• 
• 

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 (pure protein) are lyophilized.